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

UIUC - Arsenic Collection System!

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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.