User:Neil J. Parikh/Lab Notebook: Difference between revisions
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<o:LastAuthor>Neil Parikh</o:LastAuthor> | <o:LastAuthor>Neil Parikh</o:LastAuthor> | ||
<o:Revision>2</o:Revision> | <o:Revision>2</o:Revision> | ||
<o:TotalTime> | <o:TotalTime>4</o:TotalTime> | ||
<o:LastPrinted>2008-02-04T23:38:00Z</o:LastPrinted> | <o:LastPrinted>2008-02-04T23:38:00Z</o:LastPrinted> | ||
<o:Created>2008-02- | <o:Created>2008-02-06T23:47:00Z</o:Created> | ||
<o:LastSaved>2008-02- | <o:LastSaved>2008-02-06T23:47:00Z</o:LastSaved> | ||
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<o:Company>Emory Vaccine Center</o:Company> | <o:Company>Emory Vaccine Center</o:Company> | ||
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Line 1,244: | Line 1,310: | ||
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@list | @list l26 | ||
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@list l26:level1 | |||
{mso-level-number-format:roman-upper; | |||
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@list l26:level2 | |||
{mso-level-number-format:roman-upper; | |||
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margin-left:1.25in; | |||
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@list l26:level3 | |||
{mso-level-number-format:alpha-lower; | |||
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@list l26:level4 | |||
{mso-level-number-format:alpha-lower; | |||
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@list l19:level2 lfo13 | |||
{mso-level-start-at:-1073762832;} | {mso-level-start-at:-1073762832;} | ||
@list | @list l19:level3 lfo13 | ||
{mso-level-start-at:38108040;} | {mso-level-start-at:38108040;} | ||
@list | @list l19:level4 lfo13 | ||
{mso-level-start-at:-1073762928;} | {mso-level-start-at:-1073762928;} | ||
@list | @list l19:level5 lfo13 | ||
{mso-level-start-at:37978784;} | {mso-level-start-at:37978784;} | ||
@list | @list l19:level6 lfo13 | ||
{mso-level-start-at:50823780;} | {mso-level-start-at:50823780;} | ||
@list | @list l19:level7 lfo13 | ||
{mso-level-start-at:37970320;} | {mso-level-start-at:37970320;} | ||
@list | @list l19:level8 lfo13 | ||
{mso-level-start-at:268374015;} | {mso-level-start-at:268374015;} | ||
@list | @list l19:level9 lfo13 | ||
{mso-level-start-at:806748160;} | {mso-level-start-at:806748160;} | ||
ol | ol | ||
Line 1,264: | Line 1,357: | ||
ul | ul | ||
{margin-bottom:0in;} | {margin-bottom:0in;} | ||
.style1 {color: #FFFFFF} | |||
--> | --> | ||
</style> | </style> | ||
Line 1,305: | Line 1,399: | ||
<o:idmap v:ext="edit" data="1"/> | <o:idmap v:ext="edit" data="1"/> | ||
</o:shapelayout></xml><![endif]--> | </o:shapelayout></xml><![endif]--> | ||
</head> | </head> | ||
Line 1,319: | Line 1,412: | ||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | ||
padding:0in 5.4pt 0in 5.4pt'> | padding:0in 5.4pt 0in 5.4pt'> | ||
<p class=MsoTitle style='margin-right:26.1pt'>Pipetting & Making | <p class=MsoTitle style1 style='margin-right:26.1pt'>Pipetting & Making | ||
Solutions</p> | Solutions</p> </td> | ||
<td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; | |||
<td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; | |||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | ||
padding:0in 5.4pt 0in 5.4pt'> | padding:0in 5.4pt 0in 5.4pt'> | ||
<p class=ExperimentNumber>Pg. 6</p> | <p class=ExperimentNumber style1>Pg. 6</p> </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | ||
-.25in;mso-list: | -.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,340: | Line 1,430: | ||
style='color:windowtext;font-weight:normal;font-style:normal'>Pipetting<o:p></o:p></span></u></span></p> | style='color:windowtext;font-weight:normal;font-style:normal'>Pipetting<o:p></o:p></span></u></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,348: | Line 1,438: | ||
pipettes properly<u><o:p></o:p></u></span></span></p> | pipettes properly<u><o:p></o:p></u></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,356: | Line 1,446: | ||
down pipettes between solutions (with Ethanol)<u><o:p></o:p></u></span></span></p> | down pipettes between solutions (with Ethanol)<u><o:p></o:p></u></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,365: | Line 1,455: | ||
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | ||
-.25in;mso-list: | -.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | ||
</span>II.<span style='font:7.0pt "Times New Roman"'> | </span>II.<span style='font:7.0pt "Times New Roman"'> | ||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span | </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span | ||
style='color:windowtext;font-weight:normal;font-style:normal'>Solutions<o:p></o:p></span></u></span></p> | style='color:windowtext;font-weight:normal;font-style:normal'>Solutions<o:p></o:p></span></u></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,383: | Line 1,472: | ||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | ||
-9.0pt;mso-list: | -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,392: | Line 1,481: | ||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | ||
-9.0pt;mso-list: | -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,401: | Line 1,490: | ||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: | ||
-9.0pt;mso-list: | -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,419: | Line 1,508: | ||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | ||
padding:0in 5.4pt 0in 5.4pt'> | padding:0in 5.4pt 0in 5.4pt'> | ||
<p class=MsoTitle style='margin-right:26.1pt'>Plate Creation & Cell | <p class=MsoTitle style1 style='margin-right:26.1pt'>Plate Creation & Cell | ||
Culture</p> | Culture</p> </td> | ||
<td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; | <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; | ||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | ||
padding:0in 5.4pt 0in 5.4pt'> | padding:0in 5.4pt 0in 5.4pt'> | ||
<p class=ExperimentNumber>Pg. 7</p> | <p class=ExperimentNumber style1>Pg. 7</p> </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | ||
-.25in;mso-list: | -.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,442: | Line 1,528: | ||
style='color:windowtext;font-weight:normal;font-style:normal'>:<u><o:p></o:p></u></span></span></p> | style='color:windowtext;font-weight:normal;font-style:normal'>:<u><o:p></o:p></u></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,458: | Line 1,544: | ||
Bactoagar, cover with foil, autoclave for 20-30 minutes<o:p></o:p></span></span></p> | Bactoagar, cover with foil, autoclave for 20-30 minutes<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,473: | Line 1,559: | ||
font-style:normal'><o:p></o:p></span></u></span></p> | font-style:normal'><o:p></o:p></span></u></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,481: | Line 1,567: | ||
& let swirl until dissolved<u><o:p></o:p></u></span></span></p> | & let swirl until dissolved<u><o:p></o:p></u></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,490: | Line 1,576: | ||
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | <p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: | ||
-.25in;mso-list: | -.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,500: | Line 1,586: | ||
font-style:normal'>:<u><o:p></o:p></u></span></span></p> | font-style:normal'>:<u><o:p></o:p></u></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,526: | Line 1,612: | ||
with cell density measurement (using the Spec)<o:p></o:p></span></span></p> | with cell density measurement (using the Spec)<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,538: | Line 1,624: | ||
(for a miniprep); you will have to add 1mL, two times<o:p></o:p></span></span></p> | (for a miniprep); you will have to add 1mL, two times<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,550: | Line 1,636: | ||
with your Initials, Sample Name (control or experimental) and the date<o:p></o:p></span></span></p> | with your Initials, Sample Name (control or experimental) and the date<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,558: | Line 1,644: | ||
scraper in ethanol and flame lightly (2x) to sterilize<o:p></o:p></span></span></p> | scraper in ethanol and flame lightly (2x) to sterilize<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,566: | Line 1,652: | ||
cool for a few seconds<o:p></o:p></span></span></p> | cool for a few seconds<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
Line 1,575: | Line 1,661: | ||
around to ensure the bacteria breaks off into the tube<o:p></o:p></span></span></p> | around to ensure the bacteria breaks off into the tube<o:p></o:p></span></span></p> | ||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list: | <p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span | ||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | ||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | Arial;color:windowtext;font-weight:normal;font-style:normal'><span | ||
style='mso-list:Ignore'>g.<span style='font:7.0pt "Times New Roman"'> | style='mso-list:Ignore'>g.<span style='font:7.0pt "Times New Roman"'> | ||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | ||
style='color:windowtext;font-weight:normal;font-style:normal'>Set incubator at | style='color:windowtext;font-weight:normal;font-style:normal'>Set incubator at | ||
Line 1,587: | Line 1,672: | ||
font-style:normal'>C overnight and let shake gently for 12-14hrs<o:p></o:p></span></span></p> | font-style:normal'>C overnight and let shake gently for 12-14hrs<o:p></o:p></span></span></p> | ||
</ | <p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span | ||
style='color:windowtext;font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p> | |||
</ | <table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0 | ||
style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt: | |||
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'> | |||
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'> | |||
<td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt; | |||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | |||
padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoTitle style1 style='margin-right:26.1pt'>Transformation of E.coli with | |||
Recombinant DNA<o:p></o:p></p> </td> | |||
<td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; | |||
background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; | |||
padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=ExperimentNumber style1>Pg. 9<o:p></o:p></p> </td> | |||
</tr> | |||
</table> | |||
</ | <p class=MsoNormal style='margin-left:.75in'><span class=MsoIntenseEmphasis><span | ||
style='color:windowtext;font-weight:normal;font-style:normal'>*note, </span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'>the heat shock</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> to the tubes is very important for transcription to occur<o:p></o:p></span></span></p> | |||
[[ | <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | ||
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>+ | |||
Controls</u>:<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_I13522 | |||
: pTetGFP<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J45220<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group | |||
1:</u><o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_R0011<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J45219<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group | |||
2:</u><o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_I14032 | |||
/ BBa_R0051 (same)<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J06702<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group | |||
3:</u><o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J04500<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J04630 | |||
/ BBa_I13401<o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p> | |||
<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Rima | |||
& I did the transformation for Group 2:<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>I.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_I14032</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name: </span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>promoter P(Lac) IQ</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence: | |||
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>promoter</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'> </span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>(-35, -10 bases | |||
before coding)</span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>: 2007, Plate 2, Well 17K</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid: | |||
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>pSB2K3</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>II.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_R0051</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> promoter (lamda cl regulated)</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> R0051 promoter</span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location: | |||
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><i | |||
style='mso-bidi-font-style:normal'><span style='mso-fareast-font-family:Arial; | |||
mso-bidi-font-family:Arial'><span style='mso-list:Ignore'>d.<span | |||
style='font:7.0pt "Times New Roman"'> </span></span></span></i><![endif]><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'>Plasmid:</span></span><i style='mso-bidi-font-style: | |||
normal'> </i>pSB1A2<i style='mso-bidi-font-style:normal'><o:p></o:p></i></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>III.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_J06702</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> mCherry, bacterial with RBS & forward terminator</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Sequence</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'>: <o:p></o:p></span></span></p> | |||
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 | |||
style='margin-left:159.5pt;border-collapse:collapse;border:none;mso-border-alt: | |||
solid windowtext .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'> | |||
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'> | |||
<td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt; | |||
mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'>RBS<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=85 valign=top style='width:64.05pt;border:solid windowtext 1.0pt; | |||
border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: | |||
solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'>mCherry<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt; | |||
border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: | |||
solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt; | |||
border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: | |||
solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p> | |||
</td> | |||
</tr> | |||
<tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'> | |||
<td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt; | |||
border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; | |||
padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>B0034<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=85 valign=top style='width:64.05pt;border-top:none;border-left: | |||
none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | |||
mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>J06504<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=101 valign=top style='width:76.05pt;border-top:none;border-left: | |||
none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | |||
mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>B0010<o:p></o:p></span></span></p> | |||
</td> | |||
<td width=101 valign=top style='width:76.05pt;border-top:none;border-left: | |||
none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; | |||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | |||
mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>B0012<o:p></o:p></span></span></p> | |||
</td> | |||
</tr> | |||
</table> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> 2007, Plate 2, Well 11D</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'> pSB1A2</span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>IV.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Cells Used: | |||
E.coli: JM109 (Promega); Catalog #: L2001</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'><o:p> </o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>V.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>3 Plates Made:</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Control: 100μL | |||
LB / Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 1: 100μL | |||
LB/ Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span | |||
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 2: 1μL | |||
cell : 99μ H<sub>2</sub>O</span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Actual | |||
Transformation Procedure:</span></span><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'><o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level1 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>I.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>If using DNA from | |||
the registry, dilute with 9μL of H<sub>2</sub>O. If using our own DNA, use | |||
200ng. <o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>inject the water | |||
directly into the well, titerate, and then pipette it out<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>II.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Thaw 100μL | |||
competent cells on ice. (50</span></span> <span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>μL with our | |||
own/ 100</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext; | |||
font-weight:normal;font-style:normal'>μ if commercial cells used)<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>III.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Mix competent | |||
cells with 50</span></span> <span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>μ CaCl<sub>2</sub> | |||
(only if using our own cells)<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>IV.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Add 4</span></span> | |||
<span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>μL of registry DNA, or 200ng of our own DNA to the | |||
cells. <o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>V.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Put in the ice | |||
bath for about 20 minutes. <o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>VI.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Remove and | |||
immediately put in the hot water bath (42</span></span><span | |||
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; | |||
font-weight:normal;font-style:normal'>°</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>C) to heat shock for 45-60 sec.<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>VII.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Place back in the | |||
ice bath for 2 minutes. <o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>VIII.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Add 900 μL | |||
ice cold, antibiotic free LB to the cells<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>IX.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Incubate at 37</span></span><span | |||
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; | |||
font-weight:normal;font-style:normal'>°</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>C for 90 minutes in the shaking incubator.<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>X.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Dilute as desired, | |||
make the necessary controls, and plate cells using 100 μL / plate. (200 if | |||
not commercial)<o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: | |||
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> | |||
</span>XI.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>Incubate upside | |||
down (so that the label you wrote on it is face up) at 37</span></span><span | |||
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; | |||
font-weight:normal;font-style:normal'>°</span></span><span | |||
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; | |||
font-style:normal'>C for 18-24 hours, then refrigerate. <o:p></o:p></span></span></p> | |||
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span | |||
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: | |||
Arial;color:windowtext;font-weight:normal;font-style:normal'><span | |||
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> | |||
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span | |||
style='color:windowtext;font-weight:normal;font-style:normal'>The controls | |||
should not have bacteria growing on them!<o:p></o:p></span></span></p> | |||
</div> | |||
</body> | |||
</html> |
Revision as of 16:51, 6 February 2008
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</head>
<body lang=EN-US link=blue vlink=purple style='tab-interval:.5in'>
<div class=Section1>
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt: solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'> <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'> <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=MsoTitle style1 style='margin-right:26.1pt'>Pipetting & Making Solutions</p> </td> <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=ExperimentNumber style1>Pg. 6</p> </td> </tr>
</table>
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: -.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>I.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal;font-style:normal'>Pipetting<o:p></o:p></span></u></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Learned to use pipettes properly<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Make sure to wipe down pipettes between solutions (with Ethanol)<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Change tips every time you go between solutions<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: -.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>II.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal;font-style:normal'>Solutions<o:p></o:p></span></u></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Task: Make 0.3M KCl<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>i.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Salt (KCl): CAS 7447-40-7<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>ii.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>MW: 74.56g/mol<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt: -9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>iii.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>2.2368 g / 100mL<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p>
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt: solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'> <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'> <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=MsoTitle style1 style='margin-right:26.1pt'>Plate Creation & Cell Culture</p> </td> <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=ExperimentNumber style1>Pg. 7</p> </td> </tr>
</table>
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: -.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>I.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal;font-style:normal'>Steps for creating a plate</span></u></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>:<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Dissolve 25g LB in 800mL of H<sub>2</sub>O<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Bring the final volume to 1L in a 2L flask<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 15g Bactoagar, cover with foil, autoclave for 20-30 minutes<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Bring back to lab, cool to 50</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'>while stirring</span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal; font-style:normal'><o:p></o:p></span></u></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add100mg AMP & let swirl until dissolved<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Pour it into the plates (about 2-3mm deep)<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: -.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>II.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal;font-style:normal'>Cell Culture</span></u></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>:<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Use solution of LB with AMP<u><o:p></o:p></u></span></span></p>
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Ensure to keep the bags sealed to keep everything sterile <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>*Note once a bag of plates has been opened, the whole thing must be used<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>The tubes allow the solution to be aerated<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>The tubes with bacteria should look cloudy upon visual inspection—can be confirmed with cell density measurement (using the Spec)<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Sterilize the autopipette, set to 1mL (make a 2mL culture, however)<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 2mL of the LB (for a miniprep); you will have to add 1mL, two times<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Label one as the control and another as your experimental sample<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>*Label the tubes with your Initials, Sample Name (control or experimental) and the date<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Dip the culture scraper in ethanol and flame lightly (2x) to sterilize<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>e.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Let the scraper cool for a few seconds<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>f.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Scrape off a tiny bit of bacteria off of the plate and place the scraper into the tube and shake around to ensure the bacteria breaks off into the tube<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>g.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Set incubator at 37</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C overnight and let shake gently for 12-14hrs<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p>
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt: solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'> <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'> <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=MsoTitle style1 style='margin-right:26.1pt'>Transformation of E.coli with Recombinant DNA<o:p></o:p></p> </td> <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt; background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto; padding:0in 5.4pt 0in 5.4pt'> <p class=ExperimentNumber style1>Pg. 9<o:p></o:p></p> </td> </tr>
</table>
<p class=MsoNormal style='margin-left:.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>*note, </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'>the heat shock</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> to the tubes is very important for transcription to occur<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>+ Controls</u>:<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_I13522
- pTetGFP<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J45220<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group 1:</u><o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_R0011<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J45219<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group 2:</u><o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_I14032 / BBa_R0051 (same)<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J06702<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>Group 3:</u><o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J04500<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:2'> </span>BBa_J04630 / BBa_I13401<o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p>
<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Rima & I did the transformation for Group 2:<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>I.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>BBa_I14032</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>promoter P(Lac) IQ</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>promoter</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'> </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>(-35, -10 bases before coding)</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>: 2007, Plate 2, Well 17K</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>pSB2K3</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>II.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>BBa_R0051</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> promoter (lamda cl regulated)</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> R0051 promoter</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><i style='mso-bidi-font-style:normal'><span style='mso-fareast-font-family:Arial; mso-bidi-font-family:Arial'><span style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> </span></span></span></i><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'>Plasmid:</span></span><i style='mso-bidi-font-style: normal'> </i>pSB1A2<i style='mso-bidi-font-style:normal'><o:p></o:p></i></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>III.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>BBa_J06702</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> mCherry, bacterial with RBS & forward terminator</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Sequence</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'>: <o:p></o:p></span></span></p>
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
style='margin-left:159.5pt;border-collapse:collapse;border:none;mso-border-alt: solid windowtext .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'> <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'> <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt; mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'>RBS<o:p></o:p></span></span></p> </td> <td width=85 valign=top style='width:64.05pt;border:solid windowtext 1.0pt; border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'>mCherry<o:p></o:p></span></span></p> </td> <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt; border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p> </td> <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt; border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt: solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p> </td> </tr> <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'> <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt; border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>B0034<o:p></o:p></span></span></p> </td> <td width=85 valign=top style='width:64.05pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>J06504<o:p></o:p></span></span></p> </td> <td width=101 valign=top style='width:76.05pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>B0010<o:p></o:p></span></span></p> </td> <td width=101 valign=top style='width:76.05pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>B0012<o:p></o:p></span></span></p> </td> </tr>
</table>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> 2007, Plate 2, Well 11D</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> pSB1A2</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>IV.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Cells Used: E.coli: JM109 (Promega); Catalog #: L2001</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'><o:p> </o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>V.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>3 Plates Made:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Control: 100μL LB / Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 1: 100μL LB/ Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 2: 1μL cell : 99μ H<sub>2</sub>O</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><o:p> </o:p></span></span></p>
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Actual Transformation Procedure:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'><o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level1 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>I.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>If using DNA from the registry, dilute with 9μL of H<sub>2</sub>O. If using our own DNA, use 200ng. <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>inject the water directly into the well, titerate, and then pipette it out<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>II.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Thaw 100μL competent cells on ice. (50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>μL with our own/ 100</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>μ if commercial cells used)<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>III.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Mix competent cells with 50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>μ CaCl<sub>2</sub> (only if using our own cells)<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>IV.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 4</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>μL of registry DNA, or 200ng of our own DNA to the cells. <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>V.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Put in the ice bath for about 20 minutes. <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>VI.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Remove and immediately put in the hot water bath (42</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C) to heat shock for 45-60 sec.<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>VII.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Place back in the ice bath for 2 minutes. <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>VIII.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 900 μL ice cold, antibiotic free LB to the cells<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>IX.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Incubate at 37</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C for 90 minutes in the shaking incubator.<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>X.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Dilute as desired, make the necessary controls, and plate cells using 100 μL / plate. (200 if not commercial)<o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt: -.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'> </span>XI.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Incubate upside down (so that the label you wrote on it is face up) at 37</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext; font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C for 18-24 hours, then refrigerate. <o:p></o:p></span></span></p>
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family: Arial;color:windowtext;font-weight:normal;font-style:normal'><span style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'> </span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>The controls should not have bacteria growing on them!<o:p></o:p></span></span></p>
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