IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/02
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finished MG1655 genome prep (which Francis started 8/31).
MG1655 1: 187.6 ng/μL MG1655 2: 239.8 ng/μL
made dilutions to 20 ng/μL
ArsRSDM2 (for ligation with B0034)
ArsRSDM2 (for ligation with B0015)
Incubated at 37°C for 15 min, inactivated at 80°C for 20 min.
Incubate at room temp for 15 min, inactivate at 80°C for 20 min
Streaked out B0034 (ISB142) on Amp plate from glycerol stock
Streaked out B0015 (ISB147) on Amp plate and on Kan plate from glycerol stock
Transformation and Plating
100 μL cells, 3 μL ligation product, electroporate. recover in 1 mL of LB at 37°C for 1 hour
Plated 200 μL cells after 1.5 hour incubation (after 1 hour, not enough growth).
Primers: 10 micromolar ArsBF2.0 ArsBR2.0
streaked out gas vesicles (ISB157) from glycerol stock onto amp plate.