IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/27
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 UIUC - Arsenic Collection System!
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7/27/109:50 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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