Reliable genotyping of samples with very low DNA quantities using PCR

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ID: 292157
1996
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Abstract
Our purpose was to identify an experimental procedure using PCR that provides a reliable genotype at a microsatellite locus using only a few picograms of template DNA. Under these circumstances, it is possible (i) that one allele of a heterozygous individual will not be detected and (ii) that PCR-generated alleles or 'false alleles' will arise. A mathematical model has been developed to account for stochastic events when pipetting template DNA in a very dilute DNA extract and computer simulations have been performed. Laboratory experiments were also carried out using DNA extracted from a bear feces sample to determine if experimental results correlate with the mathematical model. The results of 150 typing experiments are consistent with the proposed model. Based on this model and the level of observed false alleles, an experimental procedure using the multiple tubes approach is proposed to obtain reliable genotypes with a confidence level of 99%. This multiple tubes procedure should be systematically used when genotyping nuclear loci of ancient or forensic samples, museum specimens and hair or feces of free ranging animals.
Reference Key
openalex_W1972906909 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Pierre Taberlet, Sally Griffin, Benoît Goossens, Sophie Questiau, Valérie Manceau, Nathalie Escaravage, Lisette P. Waits, Jean Bouvet
Journal Nucleic Acids Research
Year 1996
DOI
10.1093/nar/24.16.3189
URL
https://doi.org/10.1093/nar/24.16.3189
Keywords Keywords not found

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