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A simple PCR procedure for discovering microsatellites from small insert libraries
Authors:X. W. WANG  R. N. TRIGIANO  M. T. WINDHAM  R. E. DEVRIES  B. E. SCHEFFLER  T. A. RINEHART  J. M. SPIERS
Abstract: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.
Keywords:colony hybridization  flowering dogwood  genomic library  microsatellites  PCR screening  SSR enrichment
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