Genotyping of Murrah Buffalo For k-casein using Pcr -Rflp

The present study was undertaken with primary objective to genotype Murrah buffalo breeds for k-casein type by polymerase reaction-restriction fragment length polymorphism (PCR-RFLP). A 379-bp k-casein gene fragment was amplified using primers K1 (5′CAC GTC ACC CAC ACC CAC ATT TAT C-3′) AND K2 (5′-TAA TTA GCC CAT TTC GCC TTC TCT GT-3′). DNA from 20 Murrah buffaloes were extracted by phenol chloroform method, PCR was carried out in a final reaction volume of 15 μl and the reaction mixture was subjected to standard PCR protocol. PCR products (379 bp) were digested with Hinf I and Hind III (REs). Hinf I analysis yielded two fragments of 288-bp and 91 bp only for all the DNA samples hence, all the samples were typed BB. Hind III yielded two fragments of 225- and 154 bp for all DNA samples. Thus PCR-RFLP using Hinf I and Hind III revealed all the samples to be monomorphic for this locus.