Haynes Lab:Notebook/Engineering PC-TFs/2012/03/01: Difference between revisions
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* In a .5 mL tube, pipette the ligation buffer first. | |||
* Then water, DNA insert, vector, and T4 ligase. | |||
* Once completed, allow them to incubate for 10 minutes. | |||
* Meanwhile, get ice bucket and allow competent E. coli cells to thaw for five minutes. | |||
* Then follow transformation process: | |||
{| {{table}} | |||
|align="center" style="background:#f0f0f1;"|'''Transformation Process''' | |||
*Warm five 100 μg/mL Amp agar plates at 37 °C | |||
*Thaw fresh tube of DH5α Turbo cells on ice | |||
*Resuspend invisible DNA pellet (stock DNA) in 10 μL dH2O | |||
*Add 0.5 μL stock DNA to 10 μL dH2O in 0.5 mL tubes | |||
*Add 30 μL of DH5α Turbo cells to DNA + dH2O | |||
*Include #5 tube, water only, no DNA (negative control) | |||
*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 11:11, 3 April 2012
Engineering PC-TFs | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
SummaryAssembly Scheme: Number beside vector and insert is number of base pairs
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