title: A comparison of mitochondrial DNA isolation methods in frozen post-mortem human brain tissue--applications for studies of mitochondrial genetics in brain disorders.
abstract: Given that many brain disorders are characterized by mitochondrial dysfunction, there is a growing interest in investigating genetic and epigenetic variation in mitochondrial DNA (mtDNA). One major caveat for such studies is the presence of nuclear-mitochondrial pseudogenes (NUMTs), which are regions of the mitochondrial genome that have been inserted into the nuclear genome over evolution and, if not accounted for, can confound genetic studies of mtDNA. Here we provide the first systematic comparison of methods for isolating mtDNA from frozen post-mortem human brain tissue. Our data show that a commercial method from Miltenyi Biotec, which magnetically isolates mitochondria using antibodies raised against the mitochondrial import receptor subunit TOM22, gives significant mtDNA enrichment and should be considered the method of choice for mtDNA studies in frozen brain tissue.
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.
authors.forename: Matthew
authors.initials: M
authors.lastname: Devall
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.
authors.forename: Joe
authors.initials: J
authors.lastname: Burrage
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Caswell
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.
authors.forename: Matthew
authors.initials: M
authors.lastname: Johnson
authors.affiliation: Institute of Psychiatry, King's College London, London, UK.
authors.forename: Claire
authors.initials: C
authors.lastname: Troakes
authors.affiliation: Institute of Psychiatry, King's College London, London, UK.
authors.forename: Safa
authors.initials: S
authors.lastname: Al-Sarraj
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.|Institute of Psychiatry, King's College London, London, UK.
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Jeffries
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.|Institute of Psychiatry, King's College London, London, UK.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Mill
authors.affiliation: University of Exeter Medical School, University of Exeter, Devon, UK.
authors.forename: Katie
authors.initials: K
authors.lastname: Lunnon
mesh_terms: D001344:Autopsy; D001921:Brain / Q000378:metabolism*; D001927:Brain Diseases / Q000235:genetics* / Q000473:pathology; D004272:DNA, Mitochondrial / Q000235:genetics / Q000302:isolation & purification*; D019143:Evolution, Molecular; D006801:Humans; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D011544:Pseudogenes
chemical_list: D004272:DNA, Mitochondrial
keywords: DNA; brain; epigenetics; genetics; isolation; mitochondria; mtDNA; post-mortem