Development of genomic based microsatellites from Salix caprea and SSR polymorphic loci identification in Salix species
2008
Deng Yuying | Xu Lian | Zhang Bo
Chinese. 以生物素标记的(CT)15和(GT)15为探针进行杂交,借助磁珠筛选出含微卫星的Sau3A I酶切片段,构建了黄花柳微卫星富集文库,获得960个阳性克隆.随机挑选360个阳性克隆进行测序,发现含有微卫星的克隆比例达45%,最后获得的44条非同源性克隆中共含有53个微卫星位点,其中(TC/AG)n,(GA/CT)n,(CA/GT)n比例最高,占74%.随机选取其中的18个微卫星位点设计引物,用5个种个体混合DNA组成模板池,检测引物种间的多态性信息,初步筛选出5对引物在柳属种间有很好的通用性.说明微卫星富集法开发SSR标记不失为一种有效的方法.
Show more [+] Less [-]English. In present research, we constructed the microsatellites-enriched libraries of Salix caprea that contained 960 Sau 3A I-digested microsatellites positive clones identified by the approach of streptavidin magnesphere paramagnetic particles and bloted with the oligoprobes of biotinylated (CT)15 and (GT)15. Three hundred and sixty clones were randomly selected to be sequenced, and the results showed that the tested clone containing microsatelite might account for 45% and clones were identified as non-homology clones with 53 microsatellite loci, which the motif of (TC/AG)n, (GA/CT)n and (CA/GT)n repeats would be much more high, approximate 74%. The primers designed based on randomly selected 18 microsatellite loci were employed to detect the genetic polymorphism in the five Salix species, which five primer pairs of the designed primers might be suitable for the polymorphic analysis in Salix species. This research we have strongly recommended that the microsatellites-enriched procedures would be one of the efficient methods for the development of SSR in Salix.
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