Mitochondrial DNA

Methods and Protocols

Häftad, Engelska, 2024

Av Thomas J. Nicholls, Jay P. Uhler, Maria Falkenberg

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This volume compiles a comprehensive range of methods to study key aspects of mitochondrial DNA including nucleoid structure and packaging, replication, genome integrity, and disease.

Produktinformation

Utgivningsdatum2024-02-21
Mått178 x 254 x 27 mm
Vikt909 g
FormatHäftad
SpråkEngelska
SerieMethods in Molecular Biology
Antal sidor469
FörlagSpringer-Verlag New York Inc.
ISBN9781071629246

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Biovetenskap inom Naturvetenskap och teknik

Isolation of functional mitochondria and pure mtDNA from murine tissues.- Isolating mitochondria, mitoplasts, and mtDNA from cultured mammalian cells.- Coupling differential centrifugation with exonuclease treatment and size exclusion chromatography (DIFSEC) for purification of mtDNA from mammalian cells.- Isolation and Quality Control of Yeast Mitochondria.- Mitochondrial DNA isolation from plants.- Visualize the Distribution and Dynamics of Mitochondrial DNA (mtDNA) nucleoids with Multiple Labelling Strategies.- Visualization of mtDNA using FISH.- In situ analysis of mitochondrial DNA synthesis using metabolic labeling coupled to fluorescence microscopy.- Measurement of nucleoid size using STED microscopy.- How to quantify DNA compaction by TFAM with Acoustic Force Spectroscopy and Total Internal Reflection Fluorescence microscopy.- Assessing TFAM binding to human mitochondrial DNA.- Identification of proximity interactors of mammalian nucleoid proteins by BioID.- Localization of mitochondrial nucleoids by transmission electron microscopy using the transgenic expression of the mitochondrial helicase TWINKLE and APEX2.- In vitro assays of TWINKLE function.- Rolling Circle Replication and Bypass of Damaged Nucleotides.- Studying mitochondrial nucleic acid synthesis utilizing intact isolated mitochondria.- Functional assessment of mitochondrial DNA maintenance by depletion and repopulation using 2’,3’-dideoxycytidine in cultured cells.- Analysis of mitochondrial DNA replication by two-dimensional agarose gel electrophoresis.- Quantitative analysis of nucleoside triphosphate pools in mouse muscle using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry detection.- Detection of UV-Induced Deletions in Mitochondrial DNA.- Determination of the ribonucleotide content of mtDNA using alkaline gels.- 5´-end mapping in human mitochondrial DNA.- Manipulation of murine mitochondrial DNA heteroplasmy with mtZFNs.- Biolistic transformation of Chlamydomonas reinhardtii and Saccharomyces cerevisiae mitochondria.- A method for precisely identifying modifications to plant mitochondrial genomes by mitoTALENs.- Mitochondrial DNA sequencing and heteroplasmy quantification by Next Generation Sequencing.- Genomic strategies in mitochondrial diagnostics.- Mitochondrial DNA enrichment for sensitive next-generation sequencing.- Single cell analysis of mitochondrial DNA deletions.