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 = ~250 bp
Heterozygote = 169 bp and ~250 bp
Wild type = 169 bp
Separated by gel electrophoresis on a 3.0% agarose gel.