e-journal

Rapid assembly of taxonomically validated mitochondrial genomes from historical insect collections

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Polymerase chain reaction approaches for DNA sequencing from dried collection specimens of insects usually
suffer from contamination, short fragment length, and limited phylogenetic power. As an alternative, we use
shotgun metagenomic sequencing from mixed pools of DNA, from which mitochondrial sequences are filtered in
silico. We extracted DNA from a single leg of 35 species of British butterflies that had been stored in the
Natural History Museum for between 9 and 35 years. Illumina MiSeq (1/3 of flow cell) produced mitogenome
sequences of > 10 kb for ten species, of which six were (almost) complete. In addition, full or partial barcode
sequences for 31 species were recovered, which, in almost all cases, showed perfect (100%) identity to sequences
available in the BOLD database under the same taxon name. The mean length of sequence reads was 167 bp
(putative mitochondrial reads) and 153 bp (all reads). Assembly of contigs > 10 kb from these short reads was
apparently error free, except in the case of two closely-related species of Pieris that formed chimeras.
Contiguous sequences are difficult to obtain from degraded DNA with Sanger technology but the Illumina short
reads are a natural fit for dry-collection specimens. The obvious possibility for scaling up this protocol
provides exciting prospects for the large-scale indexing of natural history collections using partial or full
mitochondrial genomes.

ADDITIONAL KEYWORDS: ancientDNA – de novo assembly – DNA barcoding – Lepidoptera – museum.

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Informasi Detail

Judul Seri

Biological Journal of the Linnean Society

No. Panggil

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Penerbit

: The Linnean Society of London., 2016

Deskripsi Fisik

Biological Journal of the Linnean Society, 2016, 117, 83–95.

Bahasa

English

ISBN/ISSN

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Klasifikasi

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Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

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Subjek

BIOLOGI

Info Detail Spesifik

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Pernyataan Tanggungjawab

Wati/Agus

Versi lain/terkait

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