Determination of the rdw rat genotype by polymerase chain reaction with allele-specific primer (PCR-ASP).
The rat carrying the rdw mutation (rdw rat) is a dwarf mutant with hypothyroidism that is caused by a single G to C transversion in the thyroglobulin gene. Therefore, the development of a simple method for molecular-based genotyping of this mutation has been problematic. We have developed a rapid and simple genotyping method that provides identification of both the rdw and wild-type allele. This polymerase chain reaction with an allele-specific primer (PCR-ASP) method amplifies only the specific allele, wild or mutant type, by using 3'-terminal mismatched primers that pair only with the respective alleles. This assay should be of value for rdw colony control and rapid discrimination of rdw/rdw, rdw/+ and +/+ rats.
Masaki
Tamotsu
T
Laboratory of Animal Endocrinology and Reproduction, Faculty of Agriculture, Utsunomiya University, Tochigi, Japan.
Takamatsu
Ken
K
Umezu
Motoaki
M
D000483:Alleles; D000818:Animals; D017931:DNA Primers; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats / Q000235:genetics*; D013954:Thyroglobulin / Q000235:genetics*
D017931:DNA Primers; D013954:Thyroglobulin