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Keywords:

  • colony hybridization;
  • flowering dogwood;
  • genomic library;
  • microsatellites;
  • PCR screening;
  • SSR enrichment

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.