A simple PCR procedure for discovering microsatellites from small insert libraries
A simple PCR procedure for discovering microsatellites from small insert libraries
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DOI：
10.1111/j.1471-8286.2006.01655.x
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发表时间：
2007-07-01
期刊：
影响因子：
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通讯作者：
Spiers, J. M.
中科院分区：
文献类型：
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作者：
Wang, X. W.;Trigiano, R. N.;Spiers, J. M.
从基因组文库中发现微卫星是乏味的，因为包含插入的克隆数量少，而且由于筛选方法昂贵。本文介绍了一种筛选微卫星DNA克隆的新方法。采用两种载体标准引物和一种合成重复引物的聚合酶链反应（PCR）直接筛选菌落，而非集落杂交。在凝胶中产生强烈涂片的菌落的PCR含有所需的基序，而单一的强带表明缺乏所需的基序。这种简单的筛选方法是鉴定含微卫星菌落的一种经济有效的方法。
Microsatellite discovery from genomic libraries is tedious because of the low number of clones that contain inserts and costly because of screening methodologies. A new procedure for screening clones for microsatellite DNA is described herein. Instead of colony hybridization, a polymerase chain reaction (PCR) with two vector standard primers and one synthesized repeat primer was used to directly screen colonies. PCR of colonies that produced a strong smear in gels contained the desired motif, whereas a single strong band indicated the lack of the desired motif. This simple screening method is a cost-effective way to identify microsatellite-containing colonies.