IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/30

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9.30.10
12:20 PM

Digestion

pSB1C3 F

pSB1C3 T

ArsRSDM2

J23100B0034ArsRSDM2B0015pSB1T3

Incubated at 37°C for 15 min,inactivate at 80°C for 20 min

1:15 PM

Ligation

pSB1C3F-ArsRSDM2:

pSB1C3T-ArsRSDM2:

pSB1C3F-J2334A215T:

pSB1C3T-J2334A215T:

Incubate at room temp for 15 min, inactivate at 80°C 20 min.

2:20

Transformation

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

Plated 250 μL on Cap plates at 4:00 PM.

Also streaked out ISB169 (Cap backbone) on Cap plate.


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