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Keywords:

  • automated;
  • fluorescent;
  • genotyping;
  • high-throughput;
  • microsatellite;
  • size standard

Abstract

Current methods of automated genotyping offer many advantages over traditional gel-based approaches, including reduced handling and processing times, and increased accuracy and consistency. Unfortunately, these advances have come at a substantial cost; at present, roughly one-half of the cost of automated genotyping is due to fluorescently labelled internal size standard. Here we describe detailed methodologies for generating a highly consistent, fluorescently labelled, internal size standard using polymerase chain reaction (PCR). The methods are simple and the required reagents are inexpensive, making the in-house production of fluorescently labelled size standards a more widely accessible alternative to commercially available size standards.