Nr3c1 Genotyping: Difference between revisions
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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