User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/11

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iGEM 2011 Main project page
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Goals

  • Repeat Backbones for these PCRs


PCR

3 PCRs for TerB0015-Backbone
Oligos for 1st Terminator
Reagents μL MegaMix
H2O 24.5 74
5X Phusion Buffer HF 10 30.5
10mM dNTPs 1 each 3.1 each
Primer F 5 15.2
Primer R 5 15.2
DNA 1 3
Phusion .5 NA
3 PCR for TerB0015-Backbone
Oligos for 2nd Terminator
Reagents μL MegaMix
H2O 24.5 74
5X Phusion Buffer HF 10 30.5
10mM dNTPs 1 each 3.1 each
Primer F 5 15.2
Primer R 5 15.2
DNA 1 3
Phusion .5 NA
Cycles Gel instruct. Gel
  • 98°C 30s
  • 98°C 10s 30Cy
  • 60°C 30S 30Cy
  • 72°C 1:50 30Cy
  • 72°C 10min
  • 4°C
  • Lane2 Ladder
  • Lane3-5 Ter1-Backbone 1
  • Lane6-8 Ter2-Backbone 2



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