Isolation of microsatellites from large yellow croaker (Pseudos ciaena crocea) with PCR

Abstract

构建大黄鱼部分基因组DNA文库。以M13通用引物和根据微卫星核心序列所设计的引物,用PCR法直接对文库进行扩增,获得15个PCR阳性克隆,对阳性克隆测序。测序结果说明,6个阳性克隆中含有微卫星核心序列,用Primer 3引物设计软件对侧翼序列进行微卫星引物设计。用6对引物扩增大黄鱼基因组,PCR结果经6%聚丙烯酰胺凝胶电泳分析,其中2对引物能得到稳定的扩增。A partial large yellow croaker(Pseudosciaena crocea )genomic library was constructed.15 positive clones were isolated from screening about 500 clones of the genomic library with pcr method.The primers were (ca)7c and m13,(ag)8 and m 13,respectively.Sequencing of these clones,and 6 microsatellites were isolated.6 primers were dedigned based on unique sequences flanking each motif with the so ftware Primer3.PCR on domesticated populations was carried out with these primer s,2 of them gave steady bands.福建省自然科学基金资助项目(项目编号:B2001013

    Similar works