Difference between revisions of "TAS2R38 PCR amplification"
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# When the DNA extraction cools, vortex the tubes for 30 seconds and then set up a new 500 µL microfuge tube by adding the following: | # When the DNA extraction cools, vortex the tubes for 30 seconds and then set up a new 500 µL microfuge tube by adding the following: | ||
Revision as of 14:00, 7 August 2012
- When the DNA extraction cools, vortex the tubes for 30 seconds and then set up a new 500 µL microfuge tube by adding the following:
| Reagent | Volume |
|---|---|
| extracted DNA | 10.5 µL |
| 2X Clear GoTaq Mix | 12.5 µL |
| Forward Primer | 1.0 µL |
| Reverse Primer | 1.0 µL |
| Final Volume | 25.0 µL |
2. Run the thermocycler program called TAS2R38 with the heated lid enabled. PCR conditions:
- 5 minutes at 95˚C.
- 1 minute at 95˚C.
- 1 minute at 65˚C.
- 1 minute at 72˚C.
- repeat steps 2 - 5 29 more times.
- hold at room temperature.
- END
3. When the PCR is completed, store DNA frozen until you are ready to clean up the DNA in order to have your two alleles sequenced.
