title: Next-generation sequencing of human mitochondrial reference genomes uncovers high heteroplasmy frequency.
abstract: We describe methods for rapid sequencing of the entire human mitochondrial genome (mtgenome), which involve long-range PCR for specific amplification of the mtgenome, pyrosequencing, quantitative mapping of sequence reads to identify sequence variants and heteroplasmy, as well as de novo sequence assembly. These methods have been used to study 40 publicly available HapMap samples of European (CEU) and African (YRI) ancestry to demonstrate a sequencing error rate <5.63×10(-4), nucleotide diversity of 1.6×10(-3) for CEU and 3.7×10(-3) for YRI, patterns of sequence variation consistent with earlier studies, but a higher rate of heteroplasmy varying between 10% and 50%. These results demonstrate that next-generation sequencing technologies allow interrogation of the mitochondrial genome in greater depth than previously possible which may be of value in biology and medicine.
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
authors.forename: Maria Ximena
authors.initials: MX
authors.lastname: Sosa
authors.forename: I K Ashok
authors.initials: IK
authors.lastname: Sivakumar
authors.forename: Samantha
authors.initials: S
authors.lastname: Maragh
authors.forename: Vamsi
authors.initials: V
authors.lastname: Veeramachaneni
authors.forename: Ramesh
authors.initials: R
authors.lastname: Hariharan
authors.forename: Minothi
authors.initials: M
authors.lastname: Parulekar
authors.forename: Karin M
authors.initials: KM
authors.lastname: Fredrikson
authors.forename: Timothy T
authors.initials: TT
authors.lastname: Harkins
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Lin
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Feldman
authors.forename: Pramila
authors.initials: P
authors.lastname: Tata
authors.forename: Georg B
authors.initials: GB
authors.lastname: Ehret
authors.forename: Aravinda
authors.initials: A
authors.lastname: Chakravarti
mesh_terms: D044383:Black People / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D030541:Databases, Genetic; D014644:Genetic Variation; D054629:Genome, Mitochondrial / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D060148:HapMap Project; D006801:Humans; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods*; D044465:White People / Q000235:genetics
chemical_list: D004272:DNA, Mitochondrial