Haynes Lab:Notebook/Engineering PC-TFs/2013/03/05: Difference between revisions
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|style="background-color: # | |style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | ||
|style="background-color: # | |style="background-color: #800000" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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* BL09 | * BL09 | ||
''B)Follow digest procedure:'' | <font size=3>''B)Follow digest procedure:'' | ||
DNA 20.0 µL | DNA 20.0 µL | ||
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dH2O 5.0 uL | dH2O 5.0 uL | ||
= 30.0 uL | = 30.0 uL </font> | ||
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[[Image:gel3_5_2013.jpg| | [[Image:gel3_5_2013.jpg|200px|Stock digest 3/5/2013]] | ||
[[Image:gel3_5_2013_cutout.jpg| | [[Image:gel3_5_2013_cutout.jpg|200px|Stock digest 3/5/2013]] | ||
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<font size=4>''Transformation of hPCD, fshPCD, flyPCD''</font> | |||
<br> | |||
*Warm 100 μg/mL Amp agar plates at 37 °C | |||
*Thaw fresh tube of DH5α Turbo cells on ice | |||
*Add 30 μL of DH5α Turbo cells to DNA. | |||
*Incubate cells + DNA on ice for 5 min. | |||
*Label pre-warmed plates | |||
*Transfer cells + DNA onto agar | |||
*Add 10 - 15 sterile glass beads, shake, discard beads | |||
*Incubate plates at 37 °C overnight | |||
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Latest revision as of 22:31, 26 September 2017
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Summary
A) Get biobricks and enzymes from freezer. SpeI, PstI, XbaI The following list of parts are:
B)Follow digest procedure: DNA 20.0 µL Enzyme 1 1.0 µL Enzyme 2 1.0 µL 10x buffer 3.0 µL dH2O 5.0 uL = 30.0 uL
Biobrick Restriction Enzymes
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