IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/09/10

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9/10/10

10:15 AM

Re-Ran yesterday's PCR

Verification PCR

Ingredient Amount
10X Buffer 2 μL
10 mM dNTPs .4 μL
Template 1 μL (20 ng)
F Primer .4 μL
R Primer .4 μL
Polymerase .2 μL
H2O 15.6 μL

Samples:

  • B0034ArsRSDM2B0015pSB1C3
  • negative control DI H2O

Primers: VF2, VR, 20 micromolar

Program:

95°C 3 min
95°C 30 sec
55°C 30 sec
72°C 1 min
go to step 2 30X
72°C 3 min
hold at 4°C

2:05 PM

Gel

1%, TAE, 175 V

Lane Sample Amount
1 1 Kb Ladder 1 μL
2 b0034ArsRSDM2b0015pSB1C3 1 μL dye, 5 μL sample
3 negative control 1 μL dye, 5 μL sample
4 Tom's Sample 2 μL dye, 10 μL sample
5 Tom's Sample 2 μL dye, 10 μL sample

UIUC 9.10.10B0034ArsRSDM2B0015pSB1C3..jpg

2:45 PM

PCR Cleanup

B0034ArsRSDM2B0015pSB1C3: 23.9 ng/μL

Set up sequencing reactions:

4.5 μL DNA 6 μL VF2 (10 micromolar) 19.5 μL H2O