Department of Plant Pathology, Iowa State University, Ames, IA 50011, USA.
Biotechnol Lett. 2011 Nov;33(11):2217-23. doi: 10.1007/s10529-011-0682-9. Epub 2011 Jul 9.
Analysis of an organism's genetic diversity requires a method that gives reliable, reproducible results. Microsatellites are robust markers, however, detection of allele sizes can be difficult with some systems as well as consistency among laboratories. In this study, our two laboratories used 219 isolates of Phytophthora sojae to compare three microsatellite methods. Two capillary electrophoresis methods, the Applied Biosystems 3730 Genetic Analyzer and the CEQ 8000 Genetic Analysis system, detected an average of 2.4-fold more alleles compared to gel electrophoresis with a mean of 8.8 and 3.6 alleles per locus using capillary and gel methods, respectively. The two capillary methods were comparable, although allele sizes differed consistently by an average of 3.2 bp across isolates. Differences between capillary methods could be overcome if reference standard DNA genotypes are shared between collaborating laboratories.
分析生物体的遗传多样性需要一种能够提供可靠、可重复结果的方法。微卫星是一种强大的标记物,然而,在某些系统中检测等位基因大小以及在实验室之间保持一致性可能具有挑战性。在这项研究中,我们的两个实验室使用了 219 个大豆疫霉菌分离株来比较三种微卫星方法。两种毛细管电泳方法,即应用生物系统公司的 3730 遗传分析仪和 CEQ 8000 遗传分析系统,与凝胶电泳相比,平均检测到的等位基因数量多 2.4 倍,平均每个位点分别有 8.8 和 3.6 个等位基因。两种毛细管方法具有可比性,尽管等位基因大小在分离株之间始终相差平均 3.2bp。如果合作实验室之间共享参考标准 DNA 基因型,则可以克服毛细管方法之间的差异。