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.
Meyer
Matthias
M
Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany. mmeyer@eva.mpg.de
Stenzel
Udo
U
Myles
Sean
S
Prüfer
Kay
K
Hofreiter
Michael
M
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