Team:Arizona State/Lab/Protocols/PCR

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* Label tubes blank, 0, 1, 2, 4, 8, 10
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* Add DNA and MgCl2 to tubes according to chart:
* Add DNA and MgCl2 to tubes according to chart:
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* Add 0.5 uL Phusion polymerase to each tube.
* Add 0.5 uL Phusion polymerase to each tube.

Revision as of 20:25, 26 September 2011


Protocols: PCR


Home iGEM 2011 Home Project
Lab
Results
Notebook
Human Practices

ASU Logo.png
 

PCR:

  • Prepare master mix (for 8 tubes):
  • 10*n uL 5x Phusion buffer
  • 4*n uL 2.5 mM dNTPs
  • 2.5*n uL forward primer
  • 2.5*n uL reverse primer
  • 29.5*n uL PCR water
  • Add 48 uL master mix to each of seven tubes.
  • Label tubes blank, 0, 1, 2, 4, 8, 10
  • Add DNA and MgCl2 to tubes according to chart:

Blank 0 1 2 4 8 10
MgCl2 0 0 .5 1 2 4 5
DNA 0 1 1 1 1 1 1

  • Add 0.5 uL Phusion polymerase to each tube.
  • Place tubes in thermocycler.
  • Adjust settings as needed.

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Tempe, Arizona 85287

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