Haynes Lab:Notebook/Engineering PC-TFs/2013/03/05
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*<font size=4>''Transformation of hPCD, fshPCD, flyPCD'' | *<font size=4>''Transformation of hPCD, fshPCD, flyPCD'' | ||
| - | + | *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 23:16, 5 March 2013
 Engineering PC-TFs
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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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