Pakistan has vast population of cattle belonging to different breeds. No study on DNA typing of cattle has been conducted in Pakistan. DNA typing of cattle is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for breed characterization of cattle. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from jagular vein of two breeds of cattle (Tharparkar and Red Sindhi). DNA was extracted by Inorganic method. Primers of labeled microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these labeled microsatellite markers on 44 samples of cattle. (ienotyping analysis was performed for the PCR products of labeled microsatellite markers on agarose gel and then by the genotyper. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity and polymorphism information content (PlC) of all microsatellite markers were calculated. Average hetrozygosity, average observed homozygosity and average polymorphism infonnation content (PlC) value for all alleles was 0.60, 0.40 and 0.91 respectively. Almost all of the microsatellite markers showed significant variations in both Tharparkar and Red Sindhi breeds. Microsatellite "1NRAOO5" showed maximum variation i.e. 19 alleles and microsatellite"INRAO23" showed the least variation among all microsatellite markers i.e. 2 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Tharparkar and Red Sindhi breeds.
Results of this study lead to development of a panel of 19 microsatellite markers which can be used for breed characterization of cattle. This was a preliminary study on two cattle breeds (Tharparkar and Red Sindhi) in Pakistan. This facility can be provided on commercial basis to owners. Moreover this study can become the basis for further research investigations on cattle in Pakistan.