Lan:LE daily entries: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| (21 intermediate revisions by the same user not shown) | |||
| Line 1: | Line 1: | ||
{{Lan_head}} | |||
<center>[[Lan:Lab note books|<font face="trebuchet ms" style="color:#000080"> '''Lab note books''' </font>]]</center> | |||
Please follow this format for your entries | Please follow this format for your entries | ||
*'''Date'''(year/month/day)-Hard copy lab book reference (Initials lab book #-page) | |||
*'''Title of experiment with brief description''' | |||
*'''Experimental procedures:''' Refer to appropriate protocol and note any changes in them | |||
*'''Results:''' Post pictures with caption and appropriate identification of lanes on gels | |||
**State what are your finding and any comments | |||
<h1>2008/07/01 - '''AP06-52'''</h1> | |||
< | <h2>Expression assay on positives clones</h2> | ||
Digestion positives clone('''ND01-XX and ND01-YY''') where tested for protein expression | |||
<h3>Experimental procedures</h3> | |||
* Used O/N culture for miniprep to inoculate 10 mL 2XYTGB flasks(125 mL) to OD~0.4 | |||
* Induced with 0.1 mM IPTG (100 µL of 100 mM stock) | |||
* Incubated in shaker B at 37°C O/N | |||
* Harvested at 7:30 and lysed using lysis buffer (10X stock diluted to 1X, nothing else) | |||
<h3>Results</h3> | |||
[[Image:AP06-52.jpg]] | |||
< | <table border="1"> | ||
<caption> SDS-PAGE 15% 60 min </caption> | |||
<!-- This is the table header --> | |||
<tr> | |||
<th>Lane</th> | |||
<th>Sample</th> | |||
<th>Volume</th> | |||
</tr> | |||
<!-- These are the table rows --> | |||
<tr> | |||
<td>1</td> | |||
<td> A4-S PS</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>2</td> | |||
<td> A9-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>3</td> | |||
<td> A21-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>4</td> | |||
<td> A22-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>5</td> | |||
<td> B3-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>6</td> | |||
<td> B12-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>7</td> | |||
<td> B16-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>8</td> | |||
<td> C32-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>9</td> | |||
<td> C33-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>10</td> | |||
<td> C34-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>11</td> | |||
<td> C36-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>12</td> | |||
<td> C7-S</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>13</td> | |||
<td> C42-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>14</td> | |||
<td> C44-L</td> | |||
<td> 10 µL </td> | |||
</tr> | |||
<tr> | |||
<td>15</td> | |||
<td> PS MARKER</td> | |||
<td> 8µL </td> | |||
</tr> | |||
< | Notes: | ||
*Samples in lanes 2 and 6 had very low biomass. Apparently, #6 was for miniprep and not for protein expression. | |||
*Sample #12 was grown in SOC media (w/o glucose and pH was 8.75) | |||
<h2> Minipreps on recalcitrants clones!!</h2> | |||
*minipreps were done on clones B4-S, B6-s and B14-L | |||
Notes: Miniprep should have been done on B12-L and not B6-S | |||
Latest revision as of 06:40, 2 July 2008
Please follow this format for your entries
- Date(year/month/day)-Hard copy lab book reference (Initials lab book #-page)
- Title of experiment with brief description
- Experimental procedures: Refer to appropriate protocol and note any changes in them
- Results: Post pictures with caption and appropriate identification of lanes on gels
- State what are your finding and any comments
2008/07/01 - AP06-52
Expression assay on positives clones
Digestion positives clone(ND01-XX and ND01-YY) where tested for protein expression
Experimental procedures
- Used O/N culture for miniprep to inoculate 10 mL 2XYTGB flasks(125 mL) to OD~0.4
- Induced with 0.1 mM IPTG (100 µL of 100 mM stock)
- Incubated in shaker B at 37°C O/N
- Harvested at 7:30 and lysed using lysis buffer (10X stock diluted to 1X, nothing else)
Results
- Samples in lanes 2 and 6 had very low biomass. Apparently, #6 was for miniprep and not for protein expression.
- Sample #12 was grown in SOC media (w/o glucose and pH was 8.75)
Minipreps on recalcitrants clones!!
- minipreps were done on clones B4-S, B6-s and B14-L
Notes: Miniprep should have been done on B12-L and not B6-S
| Lane | Sample | Volume |
|---|---|---|
| 1 | A4-S PS | 10 µL |
| 2 | A9-S | 10 µL |
| 3 | A21-L | 10 µL |
| 4 | A22-L | 10 µL |
| 5 | B3-S | 10 µL |
| 6 | B12-L | 10 µL |
| 7 | B16-L | 10 µL |
| 8 | C32-S | 10 µL |
| 9 | C33-S | 10 µL |
| 10 | C34-S | 10 µL |
| 11 | C36-S | 10 µL |
| 12 | C7-S | 10 µL |
| 13 | C42-L | 10 µL |
| 14 | C44-L | 10 µL |
| 15 | PS MARKER | 8µL |
