Multiplex amplification of ancient DNA View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2006-08

AUTHORS

Holger Römpler, Paul H Dear, Johannes Krause, Matthias Meyer, Nadin Rohland, Torsten Schöneberg, Helen Spriggs, Mathias Stiller, Michael Hofreiter

ABSTRACT

This method is designed to assemble long, continuous DNA sequences using minimal amounts of fragmented ancient DNA as template. This is achieved by a two-step approach. In the first step, multiple fragments are simultaneously amplified in a single multiplex reaction. Subsequently, each of the generated fragments is amplified individually using a single primer pair, in a standard simplex (monoplex) PCR. The ability to amplify multiple fragments simultaneously in the first step allows the generation of large amounts of sequence from rare template DNA, whereas the second nested step increases specificity and decreases amplification of contaminating DNA. In contrast to current protocols using many template-consuming simplex PCRs, the method described allows amplification of several kilobases of sequence in just one reaction. It thus combines optimal template usage with a high specificity and can be performed within a day. More... »

PAGES

720-728

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nprot.2006.84

DOI

http://dx.doi.org/10.1038/nprot.2006.84

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1007310701

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/17406302


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