A method for genotype validation and primer assessment in heterozygote-deficient species, as demonstrated in the prosobranch mollusc Hydrobia ulvae.

BMC Genetics
Robert J BrownlowJohn D Fish

Abstract

In studies where microsatellite markers are employed, it is essential that the primers designed will reliably and consistently amplify target loci. In populations conforming to Hardy-Weinberg equilibrium (HWE), screening for unreliable markers often relies on the identification of heterozygote deficiencies and subsequent departures from HWE. However, since many populations naturally deviate from HWE, such as many marine invertebrates, it can be difficult to distinguish heterozygote deficiencies resulting from unreliable markers from natural processes. Thus, studies of populations that are suspected to deviate from HWE naturally would benefit from a method to validate genotype data-sets and test the reliability of the designed primers. Levels of heterozygosity are reported for the prosobranch mollusc Hydrobia ulvae (Pennant) together with a method of genotype validation and primer assessment that utilises two primer sets for each locus. Microsatellite loci presented are the first described for the species Hydrobia ulvae; the five loci presented will be of value in further study of populations of H. ulvae. We have developed a novel method of testing primer reliability in naturally heterozygote deficient populations. After the des...Continue Reading

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Citations

Mar 4, 2015·Molecular Ecology·Andia Chaves-FonnegraJose V Lopez

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Methods Mentioned

BETA
genotyping
PCR
dissection

Software Mentioned

ICROCHECKER
M ICROCHECKER
BLAST
G ENEPOP
BLASTN
CERVUS
GENEPOP
MEGA
GENEMAPPER

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