IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/22

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7/22/10
Ordered new pRpoS438F and pArsRF1 primers so they have an EcoRI site instead o XbaI.

Lawns on gas vesicle/promoter plates again

No growth on pRpoS/pArsR plates

10:25 AM

miniprepped ArsR/pSB1A2 colonies.

concentrations:

ArsR-pSB1A2 colony 1: 288.2 ng/μL

ArsR-pSB1A2 colony 2: 323.9 ng/μL

Diluted each sample to 20 ng/μL for PCR

11:00 AM

PCR

Per reaction:

18.8 μL mastermix, .2 μL F primer, .2 μL R primer, 1 μL template

Samples:

ArsR-pSB1A2 colony 1

ArsR-pSB1A2 colony 2

Negative control (no template)

Primers (20 micromolar)

VR

V2F

Program:

2:05 PM

Gel 1%, TAE 175V



3:15 PM

Site Directed Mutagenesis PCR

Modified from Quikchange kit (stratagene)

Sample:

ArsR-pSB1A2 colony 1

Primers:

ArsR SDM 1 (100 micromolar)

ArsR SDM 2 (100 micromolar)

Program:


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