A novel method of microsatellite genotyping-by-sequencing using individual combinatorial barcoding

Salla Vartia*, José L. Villanueva-Cañas, John Finarelli, Edward D. Farrell, Patrick C. Collins, Graham M. Hughes, Jeanette E L Carlsson, David T. Gauthier, Philip McGinnity, Thomas F. Cross, Richard D. Fitzgerald, Luca Mirimin, Fiona Crispie, Paul D. Cotter, Jens Carlsson

*Corresponding author for this work

Research output: Contribution to journalArticle

28 Citations (Scopus)
202 Downloads (Pure)

Abstract

This study examines the potential of next-generation sequencing based ‘genotyping-by-sequencing’ (GBS) of microsatellite loci for rapid and cost-effective genotyping in large-scale population genetic studies. The recovery of individual genotypes from large sequence pools was achieved by PCR-incorporated combinatorial barcoding using universal primers. Three experimental conditions were employed to explore the possibility of using this approach with existing and novel multiplex marker panels and weighted amplicon mixture. The GBS approach was validated against microsatellite data generated by capillary electrophoresis. GBS allows access to the underlying nucleotide sequences that can reveal homoplasy, even in large datasets and facilitates cross laboratory transfer. GBS of microsatellites, using individual combinatorial barcoding, is potentially faster and cheaper than current microsatellite approaches and offers better and more data.

Original languageEnglish
Article number150565
Number of pages9
JournalRoyal Society Open Science
Volume3
DOIs
Publication statusPublished - 20 Jan 2016

Keywords

  • Amplicon sequencing
  • Gadus morhua
  • Genotyping by sequencing
  • Next-generation sequencing
  • Ssr
  • Universal primer

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'A novel method of microsatellite genotyping-by-sequencing using individual combinatorial barcoding'. Together they form a unique fingerprint.

Cite this