Evaluating the reliability of microsatellite genotyping from low-quality DNA templates with a polynomial distribution model

Abstract

Molecular studies using trace DNA, such as from museum specimens, ancient or forensic samples and samples obtained noninvasively, often have a common problem of low quality of DNA templates. Amplification errors, such as allelic dropout and false allele, may arise during polymerase chain reaction (PCR) using such samples. A mathematical model which treats homozygotes and heterozygotes discriminately has been developed to measure sample quality and compute the confidence level of using multiple-tube approaches. We use plucked hair samples collected from 26 individual Sichuan snub-nosed monkeys (Rhinopithecus roxellana) to test the model. In this case, a confidence level of 99% can be achieved by three positive PCRs. If the sample quality is very poor and requires many PCR replicates, an alternative multiple-step genotyping method is recommended. This model enables researchers to optimize experimental protocols through pilot studies and obtain reliable genetic information using noninvasive sampling method.