Difference between revisions of "Ob/ob Genotyping"

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(wrote genotyping protocol from Ellet et al)
(No difference)

Revision as of 16:02, 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.