Full-Length Mitochondrial-DNA Sequencing on the PacBio RSII.
Conventional mitochondrial-DNA (MT DNA) sequencing approaches use Sanger sequencing of 20-40 partially overlapping PCR fragments per individual, which is a time- and resource-consuming process. We have developed a high-throughput, accurate, fast, and cost-effective human MT DNA sequencing approach. In this setup we first generate long-range PCR products for two partially overlapping 7.7 and 9.2 kb MT DNA-specific amplicons, add sample-specific barcodes, and sequence these on the PacBio RSII system to obtain full-length MT DNA sequences for genotyping/haplotyping purposes.
Department of Human Genetics; Leiden Genome Technology Center, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands. r.vossen@lumc.nl.
Rolf H A M
RH
Vossen
Department of Human Genetics; Leiden Genome Technology Center, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.
Henk P J
HP
Buermans
D004272:DNA, Mitochondrial / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004272:DNA, Mitochondrial
Haplotyping; Long read sequencing; Long-range PCR; MT-DNA; PacBio RSII; Sample multiplexing