Ob/ob Genotyping: Difference between revisions
Jump to navigation
Jump to search
wrote genotyping protocol from Ellet et al |
added categories |
||
| Line 14: | Line 14: | ||
** 58 °C 30 s | ** 58 °C 30 s | ||
** 72 °C 45 s | ** 72 °C 45 s | ||
*72 °C 7 min | *72 °C 7 min | ||
[[ Category:Genotyping ]] | |||
[[ Category:Mouse Work]] | |||
[[ Category:PCR]] | |||
[[ Category:DNA]] | |||
[[ Category:Molecular Biology]] | |||
Revision as of 16:03, 9 October 2012
Used method described in Ellet et al (http://dx.doi.org/10.1038/oby.2008.443)
Primers
- ob/ob RFLP-F: 5'- TGA GTT TGT CCA AGA TGG ACC -3'
- ob/ob RFLP-R: 5'- GCC ATC CAG GCT CTC TGG -3'
- ob/ob - WT-Lep-F: 5'- AAT GAC CTG GAG AAT CTC C -3'
PCR Reaction
The PCR program is called ob/ob and is:
- 95 °C 2 min
- 40 cycles of:
- 95 °C 30 s
- 58 °C 30 s
- 72 °C 45 s
- 72 °C 7 min