Premium
Genotyping Shorthorn cattle for generalised glycogenosis
Author(s) -
DENNIS JA,
HEALY PJ
Publication year - 2001
Publication title -
australian veterinary journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.382
H-Index - 59
eISSN - 1751-0813
pISSN - 0005-0423
DOI - 10.1111/j.1751-0813.2001.tb10897.x
Subject(s) - genotyping , amplicon , biology , genetics , allele , microbiology and biotechnology , polymerase chain reaction , exon , shorthorn , genotype , dna , gene , breed
Objective To develop a procedure for routine genotyping of Shorthorn cattle for the generalised glycogenosis allele in exon 18 of the acidic α‐glucosidase gene. Procedure Allele‐specific amplification and double mismatch amplification procedures for the discrimination of the exon 18 alleles were evaluated using leucocytes and hair roots as sources of target DNA. Results Allele‐specific amplification was effective for genotyping Shorthorn cattle at the 2454 site when purified DNA was used as target for the polymerase chain reaction. However, when the target DNA was derived from hair roots, differences in the relative yield of wild‐type and mutant amplicons were observed. The double mismatch amplification procedure was effective in genotyping all subjects, independent of the source of DNA. The unique cleavage sites for Drd I and Psh A I within exon 18 are present and absent respectively in the wildtype amplicon, and are lost and acquired, respectively, in the mutant amplicon. In addition, the Drd I and Psh A I mismatching cleavage sites incorporated into the primers serve as internal controls for Drd I and Psh A I cleavage. Conclusion The double Drd I / Psh A I mismatch amplification procedure using hair root samples as the source of DNA is a robust method for genotyping Shorthorns for generalised glycogenosis.