Mitochondrial DNA: Methods and Protocols

ISBN-13: 9781493949441 / Angielski / Miękka / 2016 / 234 str.

Matthew McKenzie

Spis treści

Part I: Detection of Mitochondrial DNA Mutations and Mitochondrial DNA Damage

1.       Comprehensive Mitochondrial Genome Analysis By Massively Parallel Sequencing

Meagan E. Palculict, Victor Wei Zhang, Lee-Jun Wong, and Jing Wang

 

2.       Analysis Of Translesion DNA Synthesis By The Mitochondrial DNA Polymerase g

William C. Copeland, Rajesh Kasiviswanathan, and Matthew J. Longley

 

3.       Quantification Of DNA Damage By Real-Time Qpcr

Wei Wang, Katja Scheffler, Ying Esbensen, And Lars Eide

 

4.       Quantitation Of Mitochondrial DNA Deletions Via Restriction Digestion/Long-Range Single Molecule

PCR

Yevgenya Kraytsberg, Xinhong Guo, Saisai Tao, Alexandra Kuznetsov, Catherine Mclean, Daniel Ehrlich,  Evan Feldman, Igor Dombrovsky,  Deye Yang, Gregory J. Cloutier, Carmen Castaneda-Sceppa, And Konstantin Khrapko

 

Part II: Visualization of Mitochondrial DNA and Mitochondrial DNA Nucleoids

5.       A Single-Cell Resolution Imaging Protocol Of Mitochondrial DNA Dynamics In Physiopathology, Mtrip, Which Also Evaluates Sublethal Cytotoxicity

Laurent Chatre, Benjamin Montagne and Miria Ricchetti

 

6.       Scalable Isolation Of Mammalian Mitochondria For Nucleic Acid And Nucleoid Analysis

Ken-Wing Lee and Daniel F. Bogenhagen

7.       Tracking Mitochondrial DNA In Situ

Anna Ligasová and Karel Koberna

 

PART II: Analysis of Mitochondrial DNA Replication, Mitochondrial DNA-Encoded Protein Translation and Mitochondrial DNA Copy Number

8.       Analysis Of Replicating Mitochondrial DNA By In Organello Labeling And Two-Dimensional Agarose Gel Electrophoresis

Ian J. Holt, Lawrence Kazak, Aurelio Reyes,  and Stuart R. Wood

 

9.       Translation And Assembly Of Radiolabelled Mitochondrial DNA-Encoded Protein Subunits From Cultured Cells And Isolated Mitochondria

Luke E. Formosa, Annette Hofer, Christin Tischner, Tina Wenz,  and Michael T. Ryan

 

10.   Analysis Of Mitochondrial DNA Copy Number And Its Regulation Through DNA Methylation Of POLGA

Xin Sun, William Lee, Vijesh Vaghjiani, and  Justin C. St. John

 

Part VI: Modification Of Mitochondrial

DNA

11.   Engineered Mtzfns For Manipulation Of Human Mitochondrial DNA Heteroplasmy

Payam A. Gammage, Lindsey Van Haute,  and Michal Minczuk

 

12.   Generation Of Xenomitochondrial Embryonic Stem Cells For The Production Of Live Xenomitochondrial Mice

Ian A. Trounce, Jessica Ackerley,  and Matthew McKenzie

 

13.   Import Of Fluorescent RNA Into Mitochondria Of Living Cells

Jaroslav Zelenka and Petr Ježek

 

Part V: Purification of Proteins Involved In Mitochondrial DNA Replication and Transcription

14.   Purification And Comparative Assay Of The Human Mitochondrial Replicative DNA Helicase

Fernando A. Rosado-Ruiz, Minyoung So, and Laurie S. Kaguni

 

15.   Expression And Purification Of Mitochondrial RNA Polymerase And Transcription Factor A From Drosophila Melanogaster

John P. Gajewski, Jamie J. Arnold, Tiina S. Salminen, Laurie S. Kaguni, and Craig E. Cameron

 

16.   Purification And Comparative Assay Of Human Mitochondrial Single-Stranded DNA-Binding Protein

Grzegorz L. Ciesielski, Fernando A. Rosado-Ruiz, and Laurie S. Kaguni

 

17.   Biolayer Interferometry: A Novel Method To Elucidate Protein- Protein And Protein- DNA Interactions In The Mitochondrial DNA Replisome

Grzegorz L. Ciesielski, Vesa P. Hytönen, and Laurie S. Kaguni

 

 

 

Blurb

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.