Difference between revisions of "Nr3c1 Genotyping"
From Bridges Lab Protocols
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==PRIMERS== | ==PRIMERS== | ||
| + | nr3c1 | ||
*Fwd: AAACAGGGTTATGCTTGGCA | *Fwd: AAACAGGGTTATGCTTGGCA | ||
*Rev: TGCCTGCTAGGCAAATGATCT | *Rev: TGCCTGCTAGGCAAATGATCT | ||
| − | + | Cre | |
| + | *Fwd: GCATT ACCGG GCAAC GAGTG ATGAG | ||
| + | *Rev: GAGTG AACGA ACCTG GTCGA AATCA GTGCG | ||
| + | |||
| + | |||
| + | Make nr3c1 primer dilution at 1uM (10uL each primer + 980uL water) and Cre at .4uM. Primers in '''Genotyping Box''' are marked with a star. | ||
| + | |||
| + | ==PCR PROTOCOL== | ||
| + | |||
| + | * 94 °C 2 min | ||
| + | * 10 cycles of: | ||
| + | ** 94 °C 20 s | ||
| + | ** 65 °C 15 s | ||
| + | ** 68 °C 10 s | ||
| + | *28 cycles of: | ||
| + | **94 °C 15 s | ||
| + | **60 °C 15 s | ||
| + | **72 °C 10 s | ||
| + | *72 °C 2 min | ||
| + | |||
| + | [[ Category:Genotyping ]] | ||
| + | [[ Category:Mouse Work]] | ||
| + | [[ Category:PCR]] | ||
| + | [[ Category:DNA]] | ||
| + | [[ Category:Molecular Biology]] | ||
Latest revision as of 19:00, 17 November 2020
PRIMERS
nr3c1
- Fwd: AAACAGGGTTATGCTTGGCA
- Rev: TGCCTGCTAGGCAAATGATCT
Cre
- Fwd: GCATT ACCGG GCAAC GAGTG ATGAG
- Rev: GAGTG AACGA ACCTG GTCGA AATCA GTGCG
Make nr3c1 primer dilution at 1uM (10uL each primer + 980uL water) and Cre at .4uM. Primers in Genotyping Box are marked with a star.
PCR PROTOCOL
- 94 °C 2 min
- 10 cycles of:
- 94 °C 20 s
- 65 °C 15 s
- 68 °C 10 s
- 28 cycles of:
- 94 °C 15 s
- 60 °C 15 s
- 72 °C 10 s
- 72 °C 2 min
