IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/12

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7/12/10
Gel

Faint bands not visible on picture for pArsRF2R1, pArsRF2R2, LamBF1R, LamB F2R

Lost pArsRF1R, pArsRF1R2 in PCR cleanup.

Eluted PCR cleanup into 2 different tubes at different volumes (50 and 20 μL):

pArsRF1R1(50ul) 5.4 ng/μL

pArsRF1R2 (50ul) 4.9 ng/μL

ArsR (50) 12 ng/μL

pArsRF1R1 (20ul) 1.8 ng/μL

pArsRF1R2 (20ul) 3.2 ng/ul

ArsR (20 ul) 4.8 ng/μL

Digestions

ArsR

pSB1A2

Incubate at 37°C for 15 minutes. Inactivate at 80°C for 20 minutes.

Gel/Gel Purification:

NB: other lanes held Tom and Amanda's samples. Ran at 175V

Purification yields:

ArsR 56.8 ng/ul

pSB1A2 8 ng/ul

Ligation

1:6 ratio of vector:insert Incubate 14°C overnight, inactivate (7/13/10 9:15AM) 80°C 20 min.


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