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Development of microsatellite genetic markers in Siberian stone pine (Pinus sibirica Du Tour) based on the de novo whole genome sequencing
Authors:M. M. Belokon,D. V. Politov,E. A. Mudrik,T. A. Polyakova,A. V. Shatokhina,Yu. S. Belokon,N. V. Oreshkova,Yu. A. Putintseva,V. V. Sharov,D. A. Kuzmin,K. V. Krutovsky
Affiliation:1.Vavilov Institute of General Genetics,Russian Academy of Sciences,Moscow,Russia;2.Russian Center of Forest Health,Federal Forestry Agency,Pushkino,Russia;3.Sukachev Institute of Forest,Siberian Branch of the Russian Academy of Sciences,Krasnoyarsk,Russia;4.Genome Research and Education Center,Siberian Federal University,Krasnoyarsk,Russia;5.Georg-August University of G?ttingen,G?ttingen,Germany;6.Texas A&M University,College Station,USA
Abstract:Siberian stone pine, Pinus sibirica Du Tour is one of the most economically and environmentally important forest-forming species of conifers in Russia. To study these forests a large number of highly polymorphic molecular genetic markers, such as microsatellite loci, are required. Prior to the new high-throughput next generation sequencing (NGS) methods, discovery of microsatellite loci and development of micro-satellite markers were very time consuming and laborious. The recently developed draft assembly of the Siberian stone pine genome, sequenced using the NGS methods, allowed us to identify a large number of microsatellite loci in the Siberian stone pine genome and to develop species-specific PCR primers for amplification and genotyping of 70 microsatellite loci. The primers were designed using contigs containing short simple sequence tandem repeats from the Siberian stone pine whole genome draft assembly. Based on the testing of primers for 70 microsatellite loci with tri-, tetra- or pentanucleotide repeats, 18 most promising, reliable and polymorphic loci were selected that can be used further as molecular genetic markers in population genetic studies of Siberian stone pine.
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