Melting curve analysis is done using a Roche Light Cycler 480.
The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX).
To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility.
Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.
Mutant Tm = 86.5°C +/- 1.0°C
Wild type Tm = 83.5°C+/- 1.0°C
Separated by gel electrophoresis on a 1.5% agarose gel.