A cost-effective approach to sequence hundreds of complete mitochondrial genomes

Joaquin C B Nunez, Marjorie F Oleksiak

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We present a cost-effective approach to sequence whole mitochondrial genomes for hundreds of individuals. Our approach uses small reaction volumes and unmodified (non-phosphorylated) barcoded adaptors to minimize reagent costs. We demonstrate our approach by sequencing 383 Fundulus sp. mitochondrial genomes (192 F. heteroclitus and 191 F. majalis). Prior to sequencing, we amplified the mitochondrial genomes using 4-5 custom-made, overlapping primer pairs, and sequencing was performed on an Illumina HiSeq 2500 platform. After removing low quality and short sequences, 2.9 million and 2.8 million reads were generated for F. heteroclitus and F. majalis respectively. Individual genomes were assembled for each species by mapping barcoded reads to a reference genome. For F. majalis, the reference genome was built de novo. On average, individual consensus sequences had high coverage: 61-fold for F. heteroclitus and 57-fold for F. majalis. The approach discussed in this paper is optimized for sequencing mitochondrial genomes on an Illumina platform. However, with the proper modifications, this approach could be easily applied to other small genomes and sequencing platforms.

Original languageEnglish (US)
Article numbere0160958
JournalPLoS One
Volume11
Issue number8
DOIs
StatePublished - Aug 1 2016

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Mitochondrial Genome
Genes
Genome
Costs and Cost Analysis
genome
Costs
Fundulidae
Fundulus
Fundulus heteroclitus
consensus sequence
Consensus Sequence
mitochondrial genome

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

A cost-effective approach to sequence hundreds of complete mitochondrial genomes. / Nunez, Joaquin C B; Oleksiak, Marjorie F.

In: PLoS One, Vol. 11, No. 8, e0160958, 01.08.2016.

Research output: Contribution to journalArticle

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