User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/08

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iGEM 2011 Main project page
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Goals

  • PCR for TerB0015-Backbone and PCR for HydEF1.

PCRs

PCR for TerB0015-Backbone
Reagents μL MegaMix
H2O 24.5 50
5X Phusion Buffer HF 10 20.1
10mM dNTPs 1 each 2 each
Primer F 5 10.2
Primer R 5 10.2
DNA 1 2
Phusion .5 NA
PCR for TerB0015-Backbone
Reagents μL MegaMix
H2O 23 46.5
5X Phusion Buffer HF 10 20.1
10mM dNTPs 1 each 2 each
Primer F 5 NA
Primer R 5 NA
DNA 1 2
DMSO 1.5 3
Phusion .5 NA
Cycles Gel instruct. Gel

Terminators

  • 98°C 30s
  • 98°C 10s 30Cy
  • 60°C 30S 30Cy
  • 72°C 1:45 30Cy
  • 72°C 10min
  • 4°C


HydEF1

  • 98°C 30s
  • 98°C 10s 30Cy
  • 60°C 30S 30Cy
  • 72°C 50 30Cy
  • 72°C 10min
  • 4°C
  • Lane1 Ladder
  • Lane2 Ter1-Backbone
  • Lane3 Ter2-Backbone
  • Lane4 HydEF 1
  • Lane5 HydEF 2

PCRs tube named as 1.0, 2.0, 3.0 and 4.0


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