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. 2017:1492:179-184.
doi: 10.1007/978-1-4939-6442-0_12.

Full-Length Mitochondrial-DNA Sequencing on the PacBio RSII

Rolf H A M Vossen  1 , Henk P J Buermans  2
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Full-Length Mitochondrial-DNA Sequencing on the PacBio RSII

Rolf H A M Vossen et al. Methods Mol Biol. 2017.

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.

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