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Mogurnda adspersa microsatellite markers: multiplexing and multi-tailed primer tagging

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A set of twelve microsatellite DNA loci were developed for the threatened Australian freshwater fish Mogurnda adspersa (Eleotridae). Primers were tailed with one of four 20-mer oligonucleotides for use in four-colour fluorescent detection and optimised for multiplex PCR. The loci were used to genotype individuals from two populations in the Pioneer River catchment of central Queensland, eastern Australia. Number of alleles per locus ranged from 2 to 33 and per locus heterozygosity ranged from 0.06 to 0.81. Successful cross-species amplification of all loci was achieved in the congener M. mogurnda. These markers will be used to estimate effective population size and to examine the relationship between flow regime and population demographic parameters.

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  • Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  • Glenn TC, Schable NA (2005) Isolating microsatellite DNA loci. Methods Enzymol 395:202–222

    Article  CAS  PubMed  Google Scholar 

  • Heegariu O, Heerema NA, Dlouhy SR, Vance GH, Vogt PH (1997) Multiplex PCR: critical parameters and step-by-step protocol. Biotechniques 23:504–511

    Google Scholar 

  • Markoulatos P, Siafakas N, Moncany M (2002) Multiplex polymerase chain reaction: a practical approach. J Clin Lab Anal 16:47–51

    Article  CAS  PubMed  Google Scholar 

  • Missiaggia A, Grattapaglia D (2006) Plant microsatellite genotyping with 4-color fluorescent detection using multiple-tailed primers. Genet Mol Res 5:72–78

    CAS  PubMed  Google Scholar 

  • Oosterhout C, Hutchinson W, Wills D, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538

    Article  Google Scholar 

  • Pusey B, Kennard M, Arthington A (2004) Freshwater fishes of North-Eastern Australia. CSIRO Publishing, Melbourne

    Google Scholar 

  • Raymond M, Rousset F (1995) genepop (Version 1.2): a population genetics software for exact tests and ecumenicism. J Hered 86(24):8–249

    Google Scholar 

  • Schuelke M (2000) An economic method for the fluorescent labelling of PCR fragments. Nat Biotechnol 18:233–234

    Article  CAS  PubMed  Google Scholar 

  • Vallone PM, Butler JM (2004) AutoDimer: a screening tool for primer-dimer and hairpin structures. Biotechniques 372:226–231

    Google Scholar 

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This work is part of a collaborative project between Griffith University and Queensland Department of Environment and Resource Management (DERM). We thank Steve Smith for refining the protocol. Bernie Cockayne and Kate Engeldow collected fish samples, Ben Cook provided M. mogurnda samples and Jake Butwell constructed laboratory equipment. Australian Animal Ethics Committee approval number for this project is ENV/08/09/AEC. Funding was provided by DERM as part of its Environmental Flows Assessment Program.

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Correspondence to Kathryn M. Real.

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Real, K.M., Schmidt, D.J. & Hughes, J.M. Mogurnda adspersa microsatellite markers: multiplexing and multi-tailed primer tagging. Conservation Genet Resour 1, 411–414 (2009).

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