Wisconsin/Protocol:PCR

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Primer Stock

First check primer concentration using spectrophotometer... then prepare stock of 500uL 10uM.

Basic Non-Sequencing PCR

  • 38.5uL H2O
  • 5uL 10x ExTaq buffer
  • 1uL 10mM dNTPs
  • 1uL Forward Primer
  • 1uL Reverse Primer
  • 1uL Template
  • 0.5uL ExTaq (add this last)
  • 2uL MgCl2 25mM

Run on Ansari PCR machine CDC-02

PCR Purification

Follow instruction in the [http://www1.qiagen.com/literature/protocols/QIAquickSpin.aspx QIAquick Spin Handbook]

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