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Title

An improved PCR method for endogenous DNA retrieval in contaminated Neandertal samples based on the use of blocking primers

AuthorsGigli, Elena ; Rasmussen, Morten; Civit, Sergi; Rosas, Antonio ; Rasilla, Marco de la; Fortea, Javier; Gilbert, M. Thomas; Willerslev, Eske; Lalueza-Fox, Carles
KeywordsNeandertals
Blocking primers
PCR
Contamination
Issue DateDec-2009
PublisherElsevier
CitationJournal of Archaeological Science 36(12): 2676-2679 (2009)
AbstractNeandertal skeletal remains are usually contaminated with modern human DNA derived from handling and washing of the specimens during excavation. Despite the fact that the distinct Neandertal haplotypes allow the design of specific primer pairs, for instance in most of the mitochondrial DNA (mtDNA) hypervariable region 1 (HVR1), the human contaminants can often outnumber the endogenous DNA, thus preventing a successful retrieval of Neandertal sequences. We have developed a novel PCR method, based on the use of blocking primers that preferentially bind to modern human contaminant DNA and block their amplification, and greatly improve the efficiency of Neandertal DNA retrieval. We tested the method in four El Sidrón Neandertal samples (two teeth and two bone fragments) with different contamination levels and taphonomic conditions, and we have been able to significantly increase the Neandertal yield from figures around 25.23% (5–69.6%) up to 90.18% (75.3–100%).
Description4 páginas, 2 tablas.
Publisher version (URL)http://dx.doi.org/10.1016/j.jas.2009.08.004
URIhttp://hdl.handle.net/10261/43703
DOI10.1016/j.jas.2009.08.004
ISSN0305-4403
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