Difference between revisions of "TAS2R38 PCR amplification"
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| − | '''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. | + | '''3.''' When the PCR is completed (~2 hours), store DNA frozen until you are ready to clean up the DNA in order to have your two alleles sequenced. |
'''4.''' Before you can send your DNA off for sequencing, you will need to [http://www.bio.davidson.edu/courses/Molbio/Protocols/Clean_Concentrate.html clean up your PCR products] | '''4.''' Before you can send your DNA off for sequencing, you will need to [http://www.bio.davidson.edu/courses/Molbio/Protocols/Clean_Concentrate.html clean up your PCR products] | ||
Revision as of 14:23, 8 August 2012
1. When the DNA extraction cools (Isolate_genomic_DNA_from_a_single_hair_follicle), 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 (~2 hours), store DNA frozen until you are ready to clean up the DNA in order to have your two alleles sequenced.
4. Before you can send your DNA off for sequencing, you will need to clean up your PCR products
Reference:
