Notes
10X AM Buffer: 67mM Tris.HCL pH8.8, 16.6mM NH4SO4, 2.0mM MgCl2, 0.17mg/ml BSA. Dissolve in 9 ml TE, pH to 8.8, then bring final volume to 10 ml with TE (NOT water!) Filter sterilize. Add 2-mercaptoethanol to 10 mM prior to use.
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.
Expected Results
Mutant = 511 bp
Heterozygote = 424 bp and 511 bp
Wild type = 424 bp
Separated by gel electrophoresis on a 1.5% agarose gel.**