IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/08/27

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8/27/10

10:00 AM

gel

Lane Sample Amount
1 1 Kb Ladder 1 μL
2 ArsB1 5 μL sample + 1 μL dye
3 ArsB2 5 μL sample + 1 μL dye
4 ArsB3 5 μL sample + 1 μL dye
5 ArsB4 5 μL sample + 1 μL dye
6 Negative control 5 μL sample + 1 μL dye

UIUC 8.27.10 arsb.jpg

11:30 AM

PCR

Ingerdient Amount Mastermix (6 reactions)
10X Buffer 2 μL 12 μL
10 mM dNTPs .4 μL 2.4 μL
Template .9 μL (18 ng) added individually to each reaction
F Primer .4 μL 2.4 μL
R Primer .4 μL 2.4 μL
Polymerase .3 μL 1.8 μL
H2O 14.6 μL 87.6 μL
DMSO 1 μL 6 μL

Samples:

  • ArsB 1
  • ArsB 2
  • ArsB 3
  • ArsB 4
  • negative control (no template)

Primers:

ArsB F 2.0, ArsB R 2.0 at 10 micromolar

Program:

95°C 3 min
95°C 30 sec
53°C 30 sec
72°C 2 min
go to step 2 32X
72°C 5 min
hold at 4°C

4:14 PM

restreaked ArsRSDM2

made 5mL overnight of ArsRSDM2-B0015 colony.