July 19, 2019  |  

Microsatellite marker discovery using single molecule real-time circular consensus sequencing on the Pacific Biosciences RS.

Authors: Grohme, Markus A and Soler, Roberto Frias and Wink, Michael and Frohme, Marcus

Microsatellite sequences are important markers for population genetics studies. In the past, the development of adequate microsatellite primers has been cumbersome. However with the advent of next-generation sequencing technologies, marker identification in genomes of non-model species has been greatly simplified. Here we describe microsatellite discovery on a Pacific Biosciences single molecule real-time sequencer. For the Greater White-fronted Goose (Anser albifrons), we identified 316 microsatellite loci in a single genome shotgun sequencing experiment. We found that the capability of handling large insert sizes and high quality circular consensus sequences provides an advantage over short read technologies for primer design. Combined with a straightforward amplification-free library preparation, PacBio sequencing is an economically viable alternative for microsatellite discovery and subsequent PCR primer design.

Journal: BioTechniques
DOI: 10.2144/000114104
Year: 2013

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