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机构地区:[1]宁波市农科院,宁波315040 [2]浙江大学农学系,杭州310029
出 处:《生物技术通报》2006年第C00期293-295,301,共4页Biotechnology Bulletin
基 金:国家转基因植物研究与产业化开发专项(J00-B-002-10);国家高技术研究发展计划(863计划;2001AA212081);国家自然科学基金(30170058);国家教育部科技研究基金(重大0114)资助项目
摘 要:应用AFLP分子标记技术,对陆地棉两个多标记基因系T582和T586厦其杂交后代F1等进行了DNA多态性分析。结果表明:在58对EcoRI/MseI引物组合中,筛选出41对引物组合具有多态性,多态性的引物组合占筛选总组合的70.69%。AFLP分子标记具有高度的多态性,非常适于基因组差异较小的(棉花)材料之间的多态性筛选。采用聚丙烯酰胺银染法显带技术,AFLP进行PCR扩增能看到30—80条DNA亮带,且检测灵敏度高,可区别只相差十几个bp甚至几个bp大小的DNA片段。但AFLP标记以显性标记占绝对优势,共显性标记比率极少,故而难以区分种质的杂合和纯合,这是它的惟一不足之处。Analysis of amplified fragment length polymorphism (AFLP) was also carried out for two parents (T582 and T586), and their F1 plants. Forty-one out of fifty-eight i. e. 70.69% of total EcoRI/MseI primers pairs were selected that could produce DNA polymorphism among T582, T586 and F1 hybrid. It's obvious that AFLP has so high rate of polymorphic primers that it is especially suitable for screening cotton varieties. Depending upon polypropylene gel electrophoresis and silver staining method, amplified products of AFLP-PCR reaction could produce 30-80 bright bands, even for those bands with less than 10bp difference. However, there was a shortage in amplification of AFLP to marker mapping because it cant distinguish between homozygous and heterozygous alleles.
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