ArticleBMC genomics2014
From cheek swabs to consensus sequences: an A to Z protocol for high-throughput DNA sequencing of complete human mitochondrial genomes.
Article in BMC genomics, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Review
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- Rapid glycoprotein evolution enables variant interactions in herpes simplex virus type 1.Virus evolution · 2025Article
- Evolutionary Dynamics of Accelerated Antiviral Resistance Development in Hypermutator Herpesvirus.Molecular biology and evolution · 2024Article
- Elucidating the role of exogenous melatonin in mitigating alkaline stress in soybeans across different growth stages: a transcriptomic and metabolomic approach.BMC plant biology · 2024Article
- Identification of non-model mammal species using the MinION DNA sequencer from Oxford Nanopore.PeerJ · 2024Article
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- Demographic, Virological Characteristics and Prognosis of Asymptomatic COVID-19 Patients in South China.Frontiers in medicine · 2022Article
- Assessment of Illumina® Human mtDNA Genome assay: workflow evaluation with development of analysis and interpretation guidelines.International journal of legal medicine · 2021Article
- Rapid and accurate species identification for ecological studies and monitoring using CRISPR-based SHERLOCK.Molecular ecology resources · 2020Article
- Detection of Innate and Artificial Mitochondrial DNA Heteroplasmy by Massively Parallel Sequencing: Considerations for Analysis.Journal of Korean medical science · 2018Article
- Ancient DNA of Phoenician remains indicates discontinuity in the settlement history of Ibiza.Scientific reports · 2018Article
- Monitored eCLIP: high accuracy mapping of RNA-protein interactions.Nucleic acids research · 2018Article
- Detection of Activating Estrogen Receptor Gene (Clinical cancer research : an official journal of the American Association for Cancer Research · 2017Article
- First complete mitochondrial genome data from ancient South American camelids - The mystery of the chilihueques from Isla Mocha (Chile).Scientific reports · 2016Article
- Changes in variation at the MHC class II DQA locus during the final demise of the woolly mammoth.Scientific reports · 2016Article
- Article
- A European Mitochondrial Haplotype Identified in Ancient Phoenician Remains from Carthage, North Africa.PloS one · 2016Article
Corrections and comments
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Authors and funding
7 authors.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundNext-generation DNA sequencing (NGS) technologies have made huge impacts in many fields of biological research, but especially in evolutionary biology. One area where NGS has shown potential is for high-throughput sequencing of complete mtDNA genomes (of humans and other animals). Despite the increasing use of NGS technologies and a better appreciation of their importance in answering biological questions, there remain significant obstacles to the successful implementation of NGS-based projects, especially for new users.
resultsHere we present an 'A to Z' protocol for obtaining complete human mitochondrial (mtDNA) genomes - from DNA extraction to consensus sequence. Although designed for use on humans, this protocol could also be used to sequence small, organellar genomes from other species, and also nuclear loci. This protocol includes DNA extraction, PCR amplification, fragmentation of PCR products, barcoding of fragments, sequencing using the 454 GS FLX platform, and a complete bioinformatics pipeline (primer removal, reference-based mapping, output of coverage plots and SNP calling).
conclusionsAll steps in this protocol are designed to be straightforward to implement, especially for researchers who are undertaking next-generation sequencing for the first time. The molecular steps are scalable to large numbers (hundreds) of individuals and all steps post-DNA extraction can be carried out in 96-well plate format. Also, the protocol has been assembled so that individual 'modules' can be swapped out to suit available resources.
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