An objective, rapid and reproducible method for scoring AFLP peak-height data that minimizes genotyping error.

Abstract:

:Amplified fragment length polymorphism (AFLP) fingerprint data are now commonly collected using DNA sequencers. AFLP genotypes are still often scored by eye from such data - a time-consuming, error-prone and subjective process. We present a semi-automated method of genotyping sequencer-collected AFLPs at predefined fragment locations (loci) within the fingerprint. Our method uses thresholds of AFLP-polymerase chain reaction-product fluorescence intensity (peak height) in order to: (i) exclude AFLP loci that are likely to contribute high rates of error to data sets, and (ii) determine the AFLP phenotype (fragment presence or absence) at the retained loci. Error rate analysis is an integral part of this process and is used to determine optimal thresholds that minimize genotyping error, while maximizing the numbers of retained loci. We show that application of this method to a large AFLP data set allows genotype calls that are rapid, objective and repeatable, facilitating the extraction of reliable genotype data for molecular ecological studies.

journal_name

Mol Ecol Resour

authors

Whitlock R,Hipperson H,Mannarelli M,Butlin RK,Burke T

doi

10.1111/j.1755-0998.2007.02073.x

subject

Has Abstract

pub_date

2008-07-01 00:00:00

pages

725-35

issue

4

eissn

1755-098X

issn

1755-0998

journal_volume

8

pub_type

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