Use of SSCP to improve the efficiency of microsatellite identification from microsatellite-enriched libraries

Authors

  • MAXINE P. PIGGOTT,

    1. Molecular Ecology Group for Marine Research, Department of Biological Sciences, Macquarie University, Sydney, New, South Wales 2109, Australia
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  • SAM C. BANKS,

    1. Molecular Ecology Group for Marine Research, Department of Biological Sciences, Macquarie University, Sydney, New, South Wales 2109, Australia
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  • LUCIANO B. BEHEREGARAY

    1. Molecular Ecology Group for Marine Research, Department of Biological Sciences, Macquarie University, Sydney, New, South Wales 2109, Australia
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Luciano B. Beheregaray, Fax: (+61 2) 9850 8245; E-mail: Luciano.Beheregaray@bio.mq.edu.au

Abstract

An inefficient aspect of marker identification from microsatellite-enriched libraries is the proportion of clones with identical sequences. This can substantially increase the number of clones that need to be sequenced in order to identify a sufficient number of microsatellite loci. We propose the use of single-stranded conformation polymorphism (SSCP) analysis to identify unique clones prior to sequencing. We used this approach prior to sequencing from microsatellite-enriched libraries for three marine invertebrate species and were able to obtain a given number of unique clone sequences for only 28% of the sequencing effort that would have been required without SSCP screening.

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