IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/08/05

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8/5/10
9:45 AM

miniprepped B0034-ArsR-B0015-pSB1C3 overnights using the promega kit. The colony 2 overnight may have spilled or something since it looked like we were missing about 3 mLs.

concentrations:

B0034-ArsR-B0015-pSB1C3 colony 1: 767.6 ng/μL

B0034-ArsR-B0015-pSB1C3 colony 2: 376.3 ng/μL

Dilutions to 20 ng/μL:

also got good growth on steak plates of the 1/2 colonies from B0034-ArsR-B0015-pSB1C3.

11:00 AM

PCR

Per Reaction: 19 μL mastermix, 1 μL template

Primers: VF, V2R 20 micromolar

Program:

Samples:   B0034-ArsR-B0015-pSB1C3 colony 1   B0034-ArsR-B0015-pSB1C3 colony 2  LamBF1R 1'  LamBF1R 2'  pArsRF1R2 1'  pArsRF1R2 2'  LamBF2R</li>  LamBF2R green</li>  pArsRF1R2 1</li>  pArsRF1R2 2</li>  Negative control (no template)</li>  Negative control (no primers, template=pArsRF1R2)</li> </ol>

3:30 PM

gel

Top:

Bottom:





Made 5 mL cultures of B0034-ArsRSDM1-B0015-pSB1C3 colonies from streak plates since we saw multiple bands in the verification gel.

4:15 PM

PCR purified ArsRSDM2 to send for sequencing tomorrow (qiagen kit). concentration=20 ng/μL.


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