Difference between revisions of "GoTaq PCR Genotyping"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (edited primer concentration) |
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==Materials== | ==Materials== | ||
*GoTaq Master Mix | *GoTaq Master Mix | ||
| − | *Primer Mix ( | + | *Primer Mix (from 100uM Primer Stocks): Add 10uL of forward and reverse primer, 20uL total, into 980uL of dH20 to make a 10uM solution. |
*Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, 14uL Nuclease-Free Water, and 1uL of template | *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. | *Gel: 0.3g Agarose to 30mL of TAE, microwave until dissolved and add 1uL of EtBr. | ||
Revision as of 14:01, 16 September 2010
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 10uM solution.
- 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.)
