GENOTYPING BOVINE-MILK PROTEINS USING ALLELE DISCRIMINATION BY PRIMERLENGTH AND AUTOMATED DNA SIZING TECHNOLOGY

A method for genotyping K-casein (A, B, E), beta-casein (A(1), A(2), A (3), A(5), B) and beta-lactoglobulin (A, B) simultaneously by the use of allele discrimination by primer length combined with automated dete ction of fragments with sequencing instrument is described. Seven diff erent mutations within the milk protein genes were analysed in order t o distinguish between the alleles examined. The samples were amplified in two separate multiplex polymerase chain reactions (PCRs), which we re then pooled and separated according to size in a single lane on the gel. By using stringent PCR conditions, we have been able to achieve allele-specific amplifications and minimize amplification of mismatche d primer for all seven mutations.