Molecular Population Genetics Group, Temasek Life Sciences Laboratory, Singapore.
Anim Genet. 2010 Apr;41(2):208-12. doi: 10.1111/j.1365-2052.2009.01973.x. Epub 2009 Sep 29.
Microsatellites are the most popular markers for parentage assignment and population genetic studies. To meet the demand for international comparability for genetic studies of Asian seabass, a standard panel of 28 microsatellites has been selected and characterized using the DNA of 24 individuals from Thailand, Malaysia, Indonesia and Australia. The average allele number of these markers was 10.82 +/- 0.71 (range: 6-19), and the expected heterozygosity averaged 0.76 +/- 0.02 (range: 0.63-1.00). All microsatellites showed Mendelian inheritance. In addition, eight standard size controls have been developed by cloning a set of microsatellite alleles into a pGEM-T vector to calibrate allele sizes determined by different laboratories, and are available upon request. Seven multiplex PCRs, each amplifying 3-5 markers, were optimized to accurately and rapidly genotype microsatellites. Parentage assignment using 10 microsatellites in two crosses (10 x 10 and 20 x 20) demonstrated a high power of these markers for revealing parent-sibling connections. This standard set of microsatellites will standardize genetic diversity studies of Asian seabass, and the multiplex PCR sets will facilitate parentage assignment.
微卫星是亲子关系鉴定和群体遗传学研究中最常用的标记。为了满足亚洲海鲈遗传研究国际可比性的需求，我们选择了 28 个微卫星作为标准标记，并对来自泰国、马来西亚、印度尼西亚和澳大利亚的 24 个个体的 DNA 进行了特征分析。这些标记的平均等位基因数为 10.82 +/- 0.71（范围：6-19），预期杂合度平均为 0.76 +/- 0.02（范围：0.63-1.00）。所有微卫星均表现出孟德尔遗传。此外，我们还通过将一组微卫星等位基因克隆到 pGEM-T 载体中，开发了 8 个标准大小的对照物，以校准不同实验室确定的等位基因大小，有需要可提供。我们还优化了 7 个多重 PCR 反应，每个反应可扩增 3-5 个标记，以准确快速地对微卫星进行基因分型。在两个杂交实验（10 x 10 和 20 x 20）中使用 10 个微卫星进行亲子关系鉴定，结果表明这些标记具有很高的揭示亲子关系的能力。这套标准的微卫星将标准化亚洲海鲈的遗传多样性研究，而多重 PCR 组将有助于亲子关系鉴定。
