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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。
Construction of partial genomic DNA library of large yellow croaker. Using M13 universal primers and primers designed according to the microsatellite core sequence, the library was directly amplified by PCR, and 15 positive clones were obtained and the positive clones were sequenced. The sequencing results showed that the six positive clones contained the microsatellite core sequence, and the primer sequences were designed by Primer 3 primer design software. Six pairs of primers were used to amplify the genome of large yellow croaker. The PCR results were analyzed by 6% polyacrylamide gel electrophoresis, of which two pairs of primers could obtain stable amplification.