User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/13

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Goal


PCR

Using .1X dilution of extraction

Reagent FinalReact MasterMix Ter1 MasterMix Ter2
H2O 24.5 50 50
5X Phusion Buffer HF 10 20.5 20.5
10mM dNTPs 1 each 2.1 each 2.1 each
Primer F 5 10.2 10.2
Primer R 5 10.2 10.2
DNA 1 2 2
Phusion .5 NA NA
Cycles GelInst Gel

98°C 10sec


30 Rep

98°C 15sec

55°C&59°C 30sec

72°C 1:50 min

72°C 10min


4°C αΩ

  • Lane1 Ladder
  • Lane2 Ter1 55°C
  • Lane2 Ter1 59°C
  • Lane2 Ter2 55°C
  • Lane2 Ter2 59°C

RepeatedPCRofTerBack1TerBack2.jpg