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This third edition is comprised of well-established protocols that are considered the gold standard in the field as well as new methodologies for mitochondrial DNA analysis. of Mitochondrial DNA: Methods and Protocols describes protocols for detecting mutations in mitochondrial DNA, techniques to assess mitochondrial DNA damage, visualization of mitochondrial DNA in situ, detection of mitochondrial DNA nucleoids within the mitochondria, methods for analyzing mitochondrial DNA replication, mitochondrial DNA-encoded protein translation and mitochondrial DNA copy number, the latest technologies for modifying the mitochondrial genome and methods for the purification of proteins involved in the replication and transcription of mitochondrial DNA. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Mitochondrial DNA: Methods and Protocols, Third Edition will be useful not only for mitochondrial researchers but also for scientists studying human diseases where mitochondrial DNA variation has been recognized as an important pathogenic factor, including cancer and neurodegeneration.
List of contents
Comprehensive Mitochondrial Genome Analysis By Massively Parallel Sequencing.- Analysis Of Translesion DNA Synthesis By The Mitochondrial DNA Polymerase g.- Quantification Of DNA Damage By Real-Time Qpcr.- Quantitation Of Mitochondrial DNA Deletions Via Restriction Digestion/Long-Range Single Molecule PCR.- A Single-Cell Resolution Imaging Protocol Of Mitochondrial DNA Dynamics In Physiopathology, Mtrip, Which Also Evaluates Sublethal Cytotoxicity.- Scalable Isolation Of Mammalian Mitochondria For Nucleic Acid And Nucleoid Analysis.- Tracking Mitochondrial DNA In Situ.- Analysis Of Replicating Mitochondrial DNA By In Organello Labeling And Two-Dimensional Agarose Gel Electrophoresis.- Translation And Assembly Of Radiolabelled Mitochondrial DNA-Encoded Protein Subunits From Cultured Cells And Isolated Mitochondria.- Analysis Of Mitochondrial DNA Copy Number And Its Regulation Through DNA Methylation Of POLGA.- Engineered Mtzfns For Manipulation Of Human Mitochondrial DNA Heteroplasm.- Generation Of Xenomitochondrial Embryonic Stem Cells For The Production Of Live Xenomitochondrial Mice.- Import Of Fluorescent RNA Into Mitochondria Of Living Cells.- Purification And Comparative Assay Of The Human Mitochondrial Replicative DNA Helicase.- Expression And Purification Of Mitochondrial RNA Polymerase And Transcription Factor A From Drosophila Melanogaster.- Purification And Comparative Assay Of Human Mitochondrial Single-Stranded DNA-Binding Protein.- Biolayer Interferometry: A Novel Method To Elucidate Protein- Protein And Protein- DNA Interactions In The Mitochondrial DNA Replisome.
Summary
This third edition is comprised of well-established protocols that are considered the gold standard in the field as well as new methodologies for mitochondrial DNA analysis. of Mitochondrial DNA: Methods and Protocols describes protocols for detecting mutations in mitochondrial DNA, techniques to assess mitochondrial DNA damage, visualization of mitochondrial DNA in situ, detection of mitochondrial DNA nucleoids within the mitochondria, methods for analyzing mitochondrial DNA replication, mitochondrial DNA-encoded protein translation and mitochondrial DNA copy number, the latest technologies for modifying the mitochondrial genome and methods for the purification of proteins involved in the replication and transcription of mitochondrial DNA. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Mitochondrial DNA: Methods and Protocols, Third Edition will be useful not only for mitochondrial researchers but also for scientists studying human diseases where mitochondrial DNA variation has been recognized as an important pathogenic factor, including cancer and neurodegeneration.
