Difference between revisions of "Ob/ob Genotyping"
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* ob/ob RFLP-R: 5'- GCC ATC CAG GCT CTC TGG -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 - 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== | ==PCR Reaction== | ||
Revision as of 16:42, 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'
- 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
