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Methods Mol Biol. 2017;1492:179-184.

Full-Length Mitochondrial-DNA Sequencing on the PacBio RSII.

Author information

1
Department of Human Genetics; Leiden Genome Technology Center, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands. r.vossen@lumc.nl.
2
Department of Human Genetics; Leiden Genome Technology Center, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.

Abstract

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.

KEYWORDS:

Haplotyping; Long read sequencing; Long-range PCR; MT-DNA; PacBio RSII; Sample multiplexing

PMID:
27822864
DOI:
10.1007/978-1-4939-6442-0_12
[Indexed for MEDLINE]

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