IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/20
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checked plates from yesterday, got lawns. inoculated 5 mL borth for each plate, incubating in 37°C room until tonight. Tomorrow will miniprep/PCR to see if insert actually in plasmid. Also made new Kan plates and will replate on these since there really shouldn't have been lawns this morning.
PCR clean up of yesterday's samples:
pRpoS 438 30.5 ng/μL
ArsR 25 ng/μL
pArsR 25.5 ng/μL
incubate at 37°C for 15 min, inactivate at 80°C for 20 min.
incubate at room temp. for 10 min, inactivate at 80°C for 20 min.
100 μL cells, 3 μL ligation product. electroporate, add 1 mL LB, recover at 37°C for 1 hour.
100 μL NEB cells, 3 μL ligation product. electroporate, add 1 mL LB, recover at 37°C for 1 hour.
200 ul on Amp plate