This assay does not work well without the use of a Hotstart Taq polymerase.
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 = ~177 bp
Heterozygote = ~177 bp and 406 bp
Wild type = 406 bp
Separated by gel electrophoresis on a 1.5% agarose gel.