GoTaq PCR Genotyping

Revision as of 16:21, 10 August 2010 by Kalefish (Talk | contribs) (Created page with '==Materials(100uM)== *GoTaq Master Mix *Primer Mix: Add 10uL of forward and reverse primer, 20uL total, into 1mL of dH20. *Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, ...')

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Revision as of 16:21, 10 August 2010 by Kalefish (Talk | contribs) (Created page with '==Materials(100uM)== *GoTaq Master Mix *Primer Mix: Add 10uL of forward and reverse primer, 20uL total, into 1mL of dH20. *Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, ...')

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Materials(100uM)

  • GoTaq Master Mix
  • Primer Mix: Add 10uL of forward and reverse primer, 20uL total, into 1mL of dH20.
  • Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, 14uL Nuclease-Free Water, and 1uL of template
  • Gel: 0.3g Agarose to 30mL of TAE, microwave until dissolved and add 1uL of EtBr.



Procedure

  • Prepare gel 30 minutes before PCR is finished.
  • Prepare Reaction Mixture, adding 13x all materials except for the template.
  • Add this mixture to each PCR tube.
  • Label and add each template to the corresponding PCR tube.
  • Run PCR under genotyping>inoki (~1 hour)
  • Load samples in gel and run on 30V (horizontally.)