IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/27

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7/27/10
9:50 AM

PCR cleanup:

LamBF1R: 5 ng/μL LamBF2R: 10 ng/μL

10:30 AM

Digestions:

LamBF1R

lamBF2R

pSB1C3 incubate at 37°C for 15 min, inactivate at 80°C for 20 min.

12:00 PM

Gel

1%, TAE, 175V



12:45 PM

Band Extraction:

pSB1C3 digest 1: 1.2 ng/μL

pSB1C3 digest 2: 1.9 ng/μL

1:20 PM

SAP

1.2 μL SAP buffer

.5 μL SAP

Incubate at 37°C for 20 min, inactivate at 65°C for 15 min.

2:25 PM

Ligations

LamBF1R-pSB1C3

LamBF2R-pSB1C3 Incubate at room temperature for 10 min, inactivate at 80°C for 20 min.

3:10 PM

Transformation and Plating

100 μL cells, 3 μL ligation product. electroporate. Add 1 mL SOC, recover at 37°C for 1 hour.

Plated 200 μL on CAP plates at 4:15 PM


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