IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/08/06

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6/27/10

9:10 AM

miniprep (promega kit):

lost about 2.5 mL of colony 2 for ArsR ligations again!

used new neutralization solution since there were chunks floating in the old solution.

concentrations:

  • B0015 pSB1AK3: 200 ng/μL
  • B0034 pSB1A2: 232.2 ng/μL
  • pSB1C3 RFP: 279.1 ng/μL
  • B0034-ArsRSDM1-B0015-pSB1C3 3 (1'): 339.2 ng/μL
  • B0034-ArsRSDM1-B0015-pSB1C3 4 (2'): 252.6 ng/μL

Glycerol stocks:

  • pARsRF1R2 1' ISB218
  • pARsRF1R2 2' ISB219

3:00 PM

Dilutions

Primers diluted to 10 micromolar

samples diluted to 20 ng/μL

Sample concentration Sample Amount H2O amount Total Vol.
VF2 100 micromolar 10 μL 90 μL 100 μL
VR 100 micromolar 10 μL 90 μL 100 μL
B0034-ArsRSDM1-B0015-pSB1C3 1 767 ng/μL .3 μL 9.7 μL 10 μL
B0034-ArsRSDM1-B0015-pSB1C3 2 376 ng/μL .53 μL 9.47 μL 10 μL
B0034-ArsRSDM1-B0015-pSB1C3 3 (1') 339.2 ng/μL .59 μL 9.41 μL 10 μL
B0034-ArsRSDM1-B0015-pSB1C3 4 (2') 252.6 ng/μL .79 μL 9.21 μL 10 μL
LamBF1R 1' 180 ng/μL 1.11 μL 8.89 μL 10 μL
LamB F1R 2' 145 ng/μL 1.38 μL 8.62 μL 10 μL
LamB F2R 68 ng/μL 2.94 μL 7.06 μL 10 μL
LamB F2R Green 285 ng/μL .7 μL 9.3 μL 10 μL
pARsR F1R2 1' 342 ng/μL .58 μL 9.42 μL 10 μL
pArsRF1R2 2' 378 ng/μL .53 μL 9.47 μL 10 μL
pArsR F1R2 1 247 ng/μL .81 μL 9.19 μL 10 μL
pArsR F1R2 2 456.2 ng/μL .44 μL 4.56 μL 10 μL




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