Targeted high-throughput sequencing of tagged nucleic acid samples.
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.
Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany. mmeyer@eva.mpg.de
Matthias
M
Meyer
Udo
U
Stenzel
Sean
S
Myles
Kay
K
Prüfer
Michael
M
Hofreiter
D004272:DNA, Mitochondrial / Q000737:chemistry; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D016324:Sequence Tagged Sites
D004272:DNA, Mitochondrial