Ob/ob Genotyping: Difference between revisions

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added other gene specific primer
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==PCR Reaction==
==PCR Reaction==
The PCR program is called '''ob/ob''' and is:
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

Bands