Haynes Lab:Notebook/Engineering PC-TFs/2013/03/05: Difference between revisions
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==Summary== | ==Summary== | ||
* | * '''Procedure''' | ||
''' A) Get biobricks and enzymes from freezer.''' | |||
SpeI, PstI, XbaI | |||
The following list of parts are: | |||
* BL01 | |||
* BL05 | |||
* BL09 | |||
<font size=3>''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 </font> | |||
''Digest Order:'' | |||
Biobrick Restriction Enzymes | |||
*1. BL01 XbaI/PstI | |||
*2. BL05 XbaI/PstI | |||
*3. BL09 XbaI/PstI | |||
[[Image:gel3_5_2013.jpg|200px|Stock digest 3/5/2013]] | |||
[[Image:gel3_5_2013_cutout.jpg|200px|Stock digest 3/5/2013]] | |||
<br> | |||
<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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Revision as of 18:23, 6 September 2013
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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
|