Single primer extension (SPEX) amplification to accurately genotype highly damaged DNA templates

Received 20 August 2007; accepted 9 October 2007.

Abstract

High levels of non-authentic sequence data can be generated by traditional PCR-based methodologies when DNA is damaged, template numbers are small and/or the target amplification size too large. We therefore present an alternate methodology based on single primer extension (SPEX) amplification; that places no pre-defined size constraints on amplification and interacts with only one of the DNA strands at the target locus.

Keywords: DNA damage, Genotyping, Low copy number DNA, SNPs

a Australian Centre for Ancient DNA, School of Earth and Environmental Sciences, University of Adelaide, Adelaide, SA 5005, Australia

b Henry Wellcome Ancient Biomolecules Centre, University of Oxford, Department of Zoology, South Parks Road, Oxford OX1 3PS, UK

c School of Animal and Microbial Sciences, University of Reading, Reading RG6 6AG, UK

d Department of Archaeology, University of Durham, South Road, Durham DH1 3L, UK

e National Institute of Toxicology and Forensic Sciences, Canary Islands delegation, 38320, Tenerife, Spain

Corresponding author. Tel.: +441457858605.

PII: S1875-1768(08)00152-2

doi:10.1016/j.fsigss.2007.10.111

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