cost-effective straightforward protocol for shotgun Illumina libraries designed to assemble complete mitogenomes from non-model species

The mitogenome is an inescapable tool in conservation biology studies. Yet, its routine sequencing may remain tricky despite next-generation sequencing technologies. An enrichment step is often necessary but not always straightforward depending on the initial DNA quality or quantity. Furthermore, th...

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Bibliographic Details
Published in:Conservation genetics resources 2015-03, Vol.7 (1), p.37-40
Main Authors: Tilak, Marie-Ka, Justy, Fabienne, Debiais-Thibaud, Mélanie, Botero-Castro, Fidel, Delsuc, Frédéric, Douzery, Emmanuel J. P
Format: Article
Language:English
Subjects:
Animal Genetics and Genomics
Biodiversity
Biomedical and Life Sciences
Chiroptera
Conservation biology
Conservation Biology/Ecology
Conservation genetics
Conserved sequence
cost effectiveness
Deoxyribonucleic acid
DNA
Ecology
Evolutionary Biology
genome
Genomes
high-throughput nucleotide sequencing
Life Sciences
Mitochondrial DNA
Next-generation sequencing
Nucleotide sequence
Plant Genetics and Genomics
polymerase chain reaction
Populations and Evolution
Species
Systematics, Phylogenetics and taxonomy
Technical Note
Tunicata
wildlife management
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Summary:The mitogenome is an inescapable tool in conservation biology studies. Yet, its routine sequencing may remain tricky despite next-generation sequencing technologies. An enrichment step is often necessary but not always straightforward depending on the initial DNA quality or quantity. Furthermore, the availability of close mitochondrial DNA reference sequences for non-model species limits the primer design for long-range PCR or bait synthesis. Here we propose an easy and cost-effective protocol without enrichment step for building and sequencing multiplexed Illumina libraries from small quantities of either high-quality or degraded genomic DNA. We validated the approach through the successful assembly of the complete mitogenome of 60 bats and 7 tunicates. Our protocol allows the sequencing and assembly of mitochondrial genomes from non-model species with sufficient coverage for applications in conservation genetics.
ISSN:1877-7252
1877-7260
DOI:10.1007/s12686-014-0338-x