Difference between revisions of "Ob/ob Genotyping"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (added other gene specific primer) |
Davebridges (Talk | contribs) m (changed program name) |
||
| Line 8: | Line 8: | ||
==PCR Reaction== | ==PCR Reaction== | ||
| − | The PCR program is called '''ob | + | The PCR program is called '''ob-ob''' and is: |
* 95 °C 2 min | * 95 °C 2 min | ||
Revision as of 15:09, 10 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'
- ob/ob - Lepob-R: 5'- GCA GAT GGA GGA GGT CTC A -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
