IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/19

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9.19.10
10:45 AM

Digestion

J23100B0034ArsRSDM2B0015pSB1T3

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

11:00 AM

PCR

Samples:   pArsR pRpoS  + control ArsR  - control H2O  

Primers: 10 micromolar   pARsRF1 2.0  pArsRR2 2.0</li>  pRpoS438 2.0</li>  pRpoSR 2.0</li>  ArsR F RFC10 EcoRI</li>  ArsR R RFC10</li> </ul>

Program: Incubate at room temperature for 15 min, inactivate at 80°C for 20 min

12:20 PM

Transformation

50 μL cells, 2 μL ligation, 10 min on ice, 45 sec at 42°C, 2 min on ice. add 1 mL LB, recover shaking at 37°C for 1 hour.

1:35 PM

Gel

TAE, 1%, 175V




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