IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/08/17

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8/17/2010
12:30PM Gel of PCR from 8/16 1% Agarose gel run at 150V for ~25min. Lane 1. 1kb ladder 2. LamB EcoRI (5uL + 1uL dye) 3. LamB EcoRI (5uL + 1uL dye) 4. (-) Control (no template)

1:00PM ArsR Re-Streak 2nd streak of ISB207 ArsR SDM2 pSB1A2

1:30PM PCR Purification PCR Purification of LamB EcoRI using Qiagen Kit

2:00PM Digestion Digesting LamB and pSB1C3 + RFP with EcoRI and SpeI

37C Incubation for 15min. 80C Inactivation for 20min.

3:30PM Ligation Ligating pSB1C3 backbone with LamB using EcoRI and SpeI ends

Room Temperature incubation for 10min. 42C for 20min -- Oops. 80C inactivation for 20min.

4:30 Transformation - Electroporation Thaw cells --> +50uL cells + 4uL Ligation mixture --> Electroporate (The machine said Arc) --> +1mL SOC medium --> 37C recovery for 1hour.

Samples: Control pUC19 Amp resistant plasmid LamB pSB1C3 ligation

Plating 300uL on pre-warmed Cap and Amp plate.


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