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 = ~300 bp (if neo is excised, no MUT band will be detected)
Heterozygote =~300 bp and 138 bp
Wild type = 138 bp (if neo is excised, a 243 bp band will be detected)
<p>Add Gel Loading Buffer (1.66ul per 12ul reaction) post PCR.</p>