Screening of microsatellite DNA sequence from the BAC library of Microtus fortis
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KeyWord:Microtus fortis;BAC library;Microsatellite;Colony hybridization;Magnetic beads
汤敏 东华大学,上海
倪丽菊 上海市实验动物质量监督检验站,上海;上海实验动物研究中心,上海
高骏 上海市农业科学院畜牧兽医研究所,上海
肖君华 东华大学,上海
胡建华 上海市实验动物质量监督检验站,上海;上海实验动物研究中心,上海
高诚 上海市实验动物质量监督检验站,上海;上海实验动物研究中心,上海
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      Objective To obtain microsatellite DNA sequence of Microtus fortis from its BAC library. MethodsTo screen the microsatellite DNA sequence with nonradioactive analysis method—colony hybridization and enrichments by magnetic beads. ResultsTwenty positive clones with the strongest signals were screened out of 136 BAC clones through hybridization of digoxin-labeled oligonucleotide (CA)20. Then these positive BAC clones were devoted to construct the subclone libraries separately. 220 microsatellite sequences were isolated from 400 positive subclones identified by PCR. According to those microsatellite sequences with more copy numbers and complete flanking sequence,74 pairs of primers were designed. As a result, 35 pairs of microsatellite primers got clear bands, of which 16 pairs had polymorphism. ConclusionsMicrosatellite DNA sequences have been screened successfully and efficiently from BAC library of Microtus fortis. The sequences and positive BAC clones may contribute to further research of their location. 
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