IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/29

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7/29/10
10:30 AM

prepared ArsR SDM1 and ArsR SDM2 samples for sequencing:

3 μL VF2 stock primer 3 μL VR stock primer 4.5 μL Template (200 ng/μL) 19.5 μL H2O

11:00

Digestions

ArsR SDM1

B0015

pSB1A2

Incubate at 37°C for 15 min, inactivate at 80°C for 20 min.

Ligations

incubate at room temperature for 10 min, inactivate at 80°C 20 min.

12:30 PM

Transformation

100 μL cells, 3 μL Ligation, electroporate, add 1 mL SOC, recover for 1 hour at 37°C

Plated 200 μL at 1:45 PM on Amp plates.

1:15 PM

PCR Per reaction: 19.2 μL mastermix, .4 μL  F Primer, .4 μL  R Primer.

Primers:

pArsR RFC10F-502 EcoRI

pArsR R1

pArsR R2

pRpoS RFC0F-438 EcoRI

pRpoS R

Samples:

pArsR F1R1

pArsR F1R2

pRpoS F-438R

Program:

4:00 PM

Gel



Made 5 mL culture of LamB colonies from yesterday. incubated at 37°C for miniprep tomorrow.


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