IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/19

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7/19/10
10:00 AM

Francis finished up genomic prep for me using the DNeasy kit. got a concentration of 98 ng/μL

diluted to 100 μL aliquots at 20ng/μL

3:20 PM

PCR Per Reaction: 19.2 μL mastermix, .4 μL F primer, .4 μL reverse primer

samples

pRpoS438

pRpoS232

ArsB

LamB F1R

LamB F2R

ArsR fusion

ArsR

pArsR F1R1

pArsR F1R2

pArsR F2R1

pArsR F2R2

Negative Control (no primers)

Primers

pRpoSF438

pRpoSF232

pRpoSR

ArsBF2

ArsBR2

ArsR F RFC10 EcoRI

ArsR R RFC10

ArsR R fusion BSAI

ArsR F fusion BSAI

pARsR F1

pArsR F2

pArsR R1

pArsR R2

LamB F1

LamB F2

LamB R

Program

6:30 PM

Gel

Top:



7:00 PM

Digestions

J23100

J23119

gas vesicles

pSB1K3

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

8:20

Ligations

J23100-gas vesicles-pSB1K3

J23119-gas vesicles-pSB1K3

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

9:15 PM

Transformations and Plating

100 μL cells, 3 μL ligation product, electroporate, add 1 mL LB, recover at 37°C for 1 hour.

Plated 200 μL cells on Kan plates at 10:15 PM.


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