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Nucleic Acids Research Advance Access originally published online on August 1, 2007
Nucleic Acids Research 2007 35(15):e97; doi:10.1093/nar/gkm566
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Nucleic Acids Research, 2007, Vol. 35, No. 15 e97
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Targeted high-throughput sequencing of tagged nucleic acid samples

Matthias Meyer*, Udo Stenzel, Sean Myles, Kay Prüfer and Michael Hofreiter

Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany

*To whom correspondence should be addressed. Tel: +49 341 3550 509, Fax: +49 341 3550 555; Email: mmeyer{at}eva.mpg.de

Received May 30, 2007. Revised June 21, 2007. Accepted July 6, 2007.

High-throughput 454 DNA sequencing technology allows much faster and more cost-effective sequencing than traditional Sanger sequencing. However, the technology imposes inherent limitations on the number of samples that can be processed in parallel. Here we introduce parallel tagged sequencing (PTS), a simple, inexpensive and flexible barcoding technique that can be used for parallel sequencing any number and type of double-stranded nucleic acid samples. We demonstrate that PTS is particularly powerful for sequencing contiguous DNA fragments such as mtDNA genomes: in theory as many as 250 mammalian mtDNA genomes can be sequenced in a single GS FLX run. PTS dramatically increases the sequencing throughput of samples in parallel and thus fully mobilizes the resources of the 454 technology for targeted sequencing.


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