title: Estimation of genotyping error rate from repeat genotyping, unintentional recaptures and known parent-offspring comparisons in 16 microsatellite loci for brown rockfish (Sebastes auriculatus).
abstract: Genotyping errors are present in almost all genetic data and can affect biological conclusions of a study, particularly for studies based on individual identification and parentage. Many statistical approaches can incorporate genotyping errors, but usually need accurate estimates of error rates. Here, we used a new microsatellite data set developed for brown rockfish (Sebastes auriculatus) to estimate genotyping error using three approaches: (i) repeat genotyping 5% of samples, (ii) comparing unintentionally recaptured individuals and (iii) Mendelian inheritance error checking for known parent-offspring pairs. In each data set, we quantified genotyping error rate per allele due to allele drop-out and false alleles. Genotyping error rate per locus revealed an average overall genotyping error rate by direct count of 0.3%, 1.5% and 1.7% (0.002, 0.007 and 0.008 per allele error rate) from replicate genotypes, known parent-offspring pairs and unintentionally recaptured individuals, respectively. By direct-count error estimates, the recapture and known parent-offspring data sets revealed an error rate four times greater than estimated using repeat genotypes. There was no evidence of correlation between error rates and locus variability for all three data sets, and errors appeared to occur randomly over loci in the repeat genotypes, but not in recaptures and parent-offspring comparisons. Furthermore, there was no correlation in locus-specific error rates between any two of the three data sets. Our data suggest that repeat genotyping may underestimate true error rates and may not estimate locus-specific error rates accurately. We therefore suggest using methods for error estimation that correspond to the overall aim of the study (e.g. known parent-offspring comparisons in parentage studies).
authors.affiliation: School of Aquatic and Fishery Sciences, University of Washington, Seattle, WA, USA. hesm@critfc.org
authors.forename: Maureen A
authors.initials: MA
authors.lastname: Hess
authors.forename: James G
authors.initials: JG
authors.lastname: Rhydderch
authors.forename: Larry L
authors.initials: LL
authors.lastname: LeClair
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Buckley
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kawase
authors.forename: Lorenz
authors.initials: L
authors.lastname: Hauser
mesh_terms: D000818:Animals; D043344:Chordata / Q000145:classification* / Q000235:genetics*; D003951:Diagnostic Errors / Q000706:statistics & numerical data; D014644:Genetic Variation; D005838:Genotype; D018895:Microsatellite Repeats; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA