Development of microsatellite markers for common bean (Phaseolus vulgaris L.) based on screening of non-enriched, small-insert genomic libraries

Genome Génome / Conseil National De Recherches Canada
Matthew W BlairNatalia Hurtado

Abstract

Microsatellite markers are useful genetic tools for a wide array of genomic analyses although their development is time-consuming and requires the identification of simple sequence repeats (SSRs) from genomic sequences. Screening of non-enriched, small-insert libraries is an effective method of SSR isolation that can give an unbiased picture of motif frequency. Here we adapt high-throughput protocols for the screening of plasmid-based libraries using robotic colony picking and filter preparation. Seven non-enriched genomic libraries from common bean genomic DNA were made by digestion with four frequently cutting restriction enzymes, double digestion with a frequently cutting restriction enzyme and a less frequently cutting restriction enzyme, or sonication. Library quality was compared and three of the small-insert libraries were selected for further analysis. Each library was plated and picked into 384-well plates that were used to create high-density filter arrays of over 18 000 clones each, which were screened with oligonucleotide probes for various SSR motifs. Positive clones were found to have low redundancy. One hundred SSR markers were developed and 80 were tested for polymorphism in a standard parental survey. These mic...Continue Reading

References

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Sep 25, 2003·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·M W BlairJ Tohme
Jan 5, 2005·Trends in Biotechnology·Rajeev K VarshneyMark E Sorrells
Apr 15, 2006·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·M W BlairS E Beebe
May 16, 2007·Genome Génome / Conseil National De Recherches Canada·Luiz Ricardo HanaiMaria Lucia Carneiro Vieira
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Sep 12, 2008·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Mathew W BlairDidier Peltier

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Citations

Apr 2, 2010·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Matthew W BlairCésar Cardona
Jun 10, 2010·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Matthew W BlairRobin Graham
Nov 30, 2010·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Matthew W BlairRobin Graham
Apr 10, 2012·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Sylvie CloutierRaja Ragupathy
Jun 22, 2012·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Matthew W Blair, Paulo Izquierdo
Aug 10, 2010·Trends in Biotechnology·Rajeev K VarshneyJean-Marcel Ribaut
Aug 26, 2016·Genetics and Molecular Biology·Maria Lucia Carneiro VieiraCarla de Freitas Munhoz
Nov 24, 2017·International Journal of Molecular Sciences·Ana M GonzálezMarta Santalla
May 1, 2012·Molecular Ecology Resources·Matthew W BlairPrem Sharma

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