Rapid Mitochondrial DNA Isolation Method for Direct Sequencing
Standard methods for mitochondrial DNA (mtDNA) extraction do not provide the level of enrichment for mtDNA sufficient for direct sequencing and must be followed by long-range-PCR amplification, which can bias the sequence results. Here, we describe a reliable method for the preparation of mtDNA-enriched samples from eukaryotic cells ready for direct sequencing. This protocol utilizes a conventional miniprep kit, in conjunction with a paramagnetic bead-based purification step.
Key wordsNext-generation sequencing Mitochondrial DNA Paramagnetic beads Solid-phase reversible immobilization Sequencing libraries
This research was supported by NIH grant P01 AG017242 (JV) and by the Ministry of Education and Science of Russian Federation grant 14.B37.21.1966 (VNP and AYM).
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