Optimisation Of Parentage Verification Methodology Using Microsatellite Markers Through Conventional PCR In Malaysian Cattle

Ernie Muneerah, Mohd Adhan, Suriaty Ramli, Mohd Hafiz Abdul Rahman,, Salleh Sheikh Ibrahim.3 & Mohd Hafizal Ahmad

Microsatellite DNA marker are a powerful tool for parentage verification. Recent studies done by Mohd-Hafiz et.al., 2012 has evaluated thirteen microsatellite markers (13) that has been recognized by the International Society of Animal Genetics (ISAG)/Food and Agricultural Organization (FAO) and also complementary marker used by international labs. To further assist with parentage analysis, this study has been done to optimize the methodology using microsatellite markers through conventional PCR for various cattle breed in Malaysia. Blood samples were collected from cattle breeds in vacutainer tube with 7.5 mg EDTA. Genomic DNA extraction was done using QIAamp DNA Blood Mini Kit (QIAGEN). Thirteen (13) microsatellite DNA regions were then amplified using conventional PCR technique and amplification reactions were done in Applied Biosystem, Veriti Model Thermocycler. PCR optimization was done using annealing temperature range from 46?C to 65?C. Amplified DNA fragments were separated using 3% MetaPhore agarose gel and were detected by Fluorosafe stain,1st Base and visualised under SYNGENE G:Box Gel Documentation and Analysis System.Besides parentage verification, this optimization result can also be used for other molecular study such as population diversity and inbreeding study.