IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/23

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7/23/10

2:45 PM

PCR purified ArsR SDM from yesterday.

Concentration after PCR purification: 20 ng/μL

3:15

DpnI Digestion

DNA 20μL (400 ng)
H2O 23.5 μL
Buffer 4 5 μL
BSA .5 μL
DpnI 1 μL

Incubate 37°C 1 hour, inactivate 80°C

4:45 PM

Transformation

100 μL NEB electrocompetent cells, 3 μL DpnI digestion product. electroporate, add 1 mL SOC, recover at 37°C for 1 hour.

100 μL our own made electrocompetent cells, 3 μL DpnI digestion product. electroporate, add 1 mL SOC, recover at 37°C for 1 hour.

5:45

Plating

plated 200 μL transformed cells onto pre-warmed Amp plates for both transformation products.