IDENTIFICATION OF OVINE AND BOVINE MICROSATELLITES IN TI-IE JAVANESE SHEEP GENOME

Abstract

Five pairs of primers flanking 2 microsatellites in the ovine genome and 3
microsatellites in the bovine genome were used to identify the corresponding microsatellites in the Javanese sheep genome as an initial step to study genetic variability within highly and lowly-prolific strains of the Javanese sheep. Each microsatellite was identified using Polymerase Chain Reaction (PCR) amplification technique with the following reagents: 50ng genomic DNA, 400 ng of each primer, 200 M dNTP’s, 1.5-3.0 n1M MgCl2, 2.5 Unit Taq Polymerase and distilled water up to 25 l. A PCR thermocycler apparatus was programmed as follows: initial denaturation at 95 °C for 3 minutes; followed by 34 cycles, each of which comprised denaturation at 95 “C, annealing at various temperatures, DNA extension at 72 “C for I5 seconds respectively; then finalized by another extension step at 72 "C for 5 minutes. PCR products were electrophoretically separated in 1% normal agarose gel and visualized under Ultra Violet (UV) transillumination. Primer OarCP79 derived from the ovine microsatellite flanking regions and primers BL25 and BM2320 derived from the bovine microsatellite flanking regions successfully
amplified single DNA segments in the Javanese sheep genome; the other ovine-derived primer OarFCB20 as well as bovine-derived primer BL37 are still being examined to amplify any DNA segment from the Javanese genome. This preliminary result indicated that primers flanking microsatellites isolated from both species and probably fi'om other species are potential to generate a number of DNA markers for genetic studies in the Javanese sheep.