| 曼氏无针乌贼GT微卫星位点的筛选 |
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| 引用本文： | 吴璋,张晓菊,蒋霞敏,薛良义.曼氏无针乌贼GT微卫星位点的筛选[J].生物技术通报,2011(3). |
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| 作者姓名： | 吴璋 张晓菊 蒋霞敏 薛良义 |
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| 作者单位： | 宁波大学生命科学与生物工程学院,宁波,315211 |
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| 基金项目： | 浙江省重大专项(2007C12061); 国家农业成果转化项目(2009GB2C220415) |
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| 摘 要： | 采用链霉亲和素包被的磁珠富集法筛选曼氏无针乌贼(Sepiella maindroni)微卫星位点。试验样品来自舟山六横岛,提取4个样品的DNA混合成DNA pool,用限制性内切酶Sau 3A I酶切。接上接头后构建基因组PCR文库,用生物素标记的(GT)15探针筛选。将筛选获得目的片段进行PCR扩增,连接pMD18-T载体,转入DH5α感受态大肠杆菌里,扩大培养后PCR筛选阳性克隆。总共选取278个克隆,对120个经过检测含有插入片段的克隆进行测序,发现102个克隆含有微卫星序列,阳性克隆比率为85%。除去重复测序和侧翼链不足的序列,可以设计引物的微卫星序列有64条。
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| 关 键 词： | 曼氏无针乌贼 微卫星 磁珠 |
Isolation of GT Microsatellite Loci from Sepiella maindroni |
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| Wu Zhang,Zhang Xiaoju,Jiang Xiamin,Xue Liangyi.Isolation of GT Microsatellite Loci from Sepiella maindroni[J].Biotechnology Bulletin,2011(3). |
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| Authors: | Wu Zhang Zhang Xiaoju Jiang Xiamin Xue Liangyi |
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| Institution: | Wu Zhang Zhang Xiaoju Jiang Xiamin Xue Liangyi(Faculty of Life Science and Biotechnology,Ningbo University,Ningbo 315211) |
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| Abstract: | Microsatellites are now widely applied in genetic studies even though they are relatively difficult to obtain.In this experiment,biotinylated magnetic beads are used to enrich microsatellite loci of Sepiella maindroni.DNA pool,which is mixed from genetic DNA of 4 different individuals derived from Liuheng island of Zhoushan,was digested by restriction endonuclease Sau 3A I.After ligation of specific adaptor,digested fragments were amplified via PCR to construct a DNA library,from which microsatellites were ... |
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| Keywords: | Sepiella maindroni Microsatellite Magnetic beads |
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