GoTaq PCR Genotyping: Difference between revisions
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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 1uM solution. | ||
*Reaction Mixture: 25uL GoTaq Mix, 10uL of Primer Mix, | *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. | *Gel: 0.3g Agarose to 30mL of TAE, microwave until dissolved and add 1uL of EtBr. | ||
| Line 12: | Line 12: | ||
*Add this mixture to each PCR tube. | *Add this mixture to each PCR tube. | ||
*Label and add each template to the corresponding PCR tube. | *Label and add each template to the corresponding PCR tube. | ||
*Run PCR under | *Run PCR under appropriate [[Genotyping Program]] | ||
*Load samples in gel and run on | *Load samples in gel and run on 130V (horizontally.) | ||
[[Category:Mouse Work]] | |||
[[Category:PCR]] | |||
[[Category:Genotyping]] | |||
Latest revision as of 14:50, 6 December 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 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.)