Difference between revisions of "GoTaq PCR Genotyping"
From Bridges Lab Protocols
(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 | + | ==Materials== |
*GoTaq Master Mix | *GoTaq Master Mix | ||
| − | *Primer Mix: Add 10uL of forward and reverse primer, 20uL total, into 1mL of dH20. | + | *Primer Mix (100uL Primers): 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 | *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 16:21, 10 August 2010
Materials
- GoTaq Master Mix
- Primer Mix (100uL Primers): 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.)
