Dept. of Animal Biotechnology, Veterinary College, Anand Agricultural University, Anand, Gujarat, India-388001.
J Anim Sci. 2012 Dec;90(13):4723-8. doi: 10.2527/jas.2011-4353. Epub 2012 Jul 31.
The present study was undertaken to construct a multiplex microsatellite panel for parentage testing in Mehsana buffalo (Bubalus bubalis). The study was based on a total of 212 Mehsana buffalos (100 dams, 100 daughters, and 12 sires). Genomic DNA was extracted from blood and semen samples. A panel of 10 microsatellite markers (CSSM61, ILSTS29, ILSTS17, ILSTS28, CSSM57, CSSM22, ILSTS61, CSSM8, ETH152, and ILSTS11) was amplified in a single multiplex reaction and analyzed by capillary electrophoresis on an automated DNA sequencer. The expected heterozygosity ranged from 0.642 to 0.833 (mean 0.762). The total exclusion probability using 10 microsatellite loci with 1 known parent was 0.993. Seven out of 10 microsatellite loci revealed relatively high polymorphic information content (>0.7). Eighty-one daughters out of 100 daughters qualified by compatibility according to Mendelism. The results suggest that multiplex microsatellite panel is a fast, robust, reliable, and economic tool to verify the parentage as well as to assign the putative sire to daughters under progeny testing with very high accuracy and hence can be used in routine parentage testing.
本研究旨在构建一个用于梅赫萨纳水牛（Bubalus bubalis）亲子鉴定的多重微卫星面板。该研究基于总共 212 头梅赫萨纳水牛（100 头母畜、100 头女儿和 12 头公牛）。从血液和精液样本中提取基因组 DNA。在单个多重反应中扩增了 10 个微卫星标记（CSSM61、ILSTS29、ILSTS17、ILSTS28、CSSM57、CSSM22、ILSTS61、CSSM8、ETH152 和 ILSTS11），并在自动 DNA 测序仪上通过毛细管电泳进行分析。预期杂合度范围为 0.642 至 0.833（平均值为 0.762）。使用 10 个微卫星标记（1 个已知亲代）的总排除概率为 0.993。10 个微卫星标记中有 7 个显示出相对较高的多态信息含量（>0.7）。根据孟德尔定律，100 头女儿中有 81 头符合兼容性要求。结果表明，多重微卫星面板是一种快速、稳健、可靠和经济的工具，可用于验证亲子关系以及根据后代测试非常准确地将假定的种公牛分配给女儿，因此可用于常规亲子关系测试。
