GoTaq PCR Genotyping
From Bridges Lab Protocols
Materials
- GoTaq Master Mix
- Primer Mix (from 100uM Primer Stocks): Add 10uL of forward and reverse primer, 20uL total, into 980uL of dH20 to make a 1uM solution.
- Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, 15uL of template. If doing several samples make a master mix of primer mix + template (25 uL GOTaq + 10 uL Primer Mix)x # of Samples + 1
- 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 appropriate Genotyping Program
- Load samples in gel and run on 130V (horizontally.)