IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/02

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9/2/10
12:30 PM

finished MG1655 genome prep (which Francis started 8/31).

MG1655 1: 187.6 ng/μL MG1655 2: 239.8 ng/μL

made dilutions to 20 ng/μL

1:30 PM

Digestions

ArsRSDM2 (for ligation with B0034)

ArsRSDM2 (for ligation with B0015)

B0015

pSB1A2

pSB1K3

Incubated at 37°C for 15 min, inactivated at 80°C for 20 min.

2:15 PM

Ligations

ArsRSDM2-B0034-pSB1K3

ArsRSDM2-B0015-pSB1K3

ArsRSDM2-B0015-pSB1A2

Incubate at room temp for 15 min, inactivate at 80°C for 20 min

2:10 PM

Streaked out B0034 (ISB142) on Amp plate from glycerol stock

Streaked out B0015 (ISB147) on Amp plate and on Kan plate from glycerol stock

3:20 PM

Transformation and Plating

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

Plated 200 μL cells after 1.5 hour incubation (after 1 hour, not enough growth).

4:10 PM

'PCR

Samples:

  ArsB 1 ArsB 2   ArsB 3  ArsB 4  Negative control (no template) 

Primers: 10 micromolar ArsBF2.0 ArsBR2.0

Program:

°C

5:30 PM

streaked out gas vesicles (ISB157) from glycerol stock onto amp plate.


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