Main description:

The aim of this volume is to provide a broad overview of single-molecule approaches applied to biomolecules. Chapters in Single Molecule Techniques: Methods and Protocols, Second Edition detail the most widely used single-molecule techniques, such as DNA, DNA-binding proteins, motor proteins, and are becoming commonplace in molecular biophysics, biochemistry, and molecular and cell biology. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Single Molecule Analysis: Methods and Protocols, Second Edition aims to be useful to many researchers, inspire them and help them to go single molecule.

Contents:

Part I

1. Introduction to Optical Tweezers: Background, System Designs, and Commercial Solutions
Joost van Mameren, Gijs J. L. Wuite, and Iddo Heller

2. RNA Unzipping and Force Measurements with a Dual Optical Trap
Laurent Geffroy, Pierre Mangeol, Thierry Bizebard, and Ulrich Bockelmann

3. Protein Tethering for Folding Studies
Fatemeh Moayed, Roeland J. van Wijk, David P. Minde, and Sander J. Tans

4. Combining Structure-Function and Single-Molecule Studies on Cytoplasmic Dynein
Lu Rao, Maren Hulsemann, and Arne Gennerich

5. A Brief Introduction to Single-Molecule Fluorescence Methods
Siet M. J. L. van den Wildenberg, Bram Prevo, and Erwin J. G. Peterman

PART II

6. Fluorescent Labeling of Proteins
Mauro Modesti

7. Single-Molecule Imaging of Escherichia coli Trans-Membrane Proteins
Aravindan Varadarajan, Felix Oswald, and Yves J. M. Bollen

8. Single-Molecule Fluorescence Microscopy in Living Caenorhabditis elegans
Jaap van Krugten and Erwin J. G. Peterman

9. Purification and Application of a Small Actin Probe for Single-Molecule Localization Microscopy
Roderick P. Tas, Trusanne G. A. A. Bos, and Lukas C. Kapitein

10. Fluorescence Microscopy of Nanochannel-Confined DNA
Fredrik Westerlund, Fredrik Persson, Joachim Fritsche, Jason P. Beech, and Jonas O. Tegenfeldt

11. Use of Single Molecule Fluorescence Polarization Microscopy to Study Protein Conformation and Dynamics of Kinesin--Microtubule Complexes
Matthieu P. M. H. Benoit and Hernando Sosa

12. Single Molecule FRET Analysis of DNA Binding Proteins
Kathy R. Chaurasiya and Remus T. Dame

PART III

13. Atomic Force Microscopy: An Introduction
Melissa C. Piontek and Wouter H. Roos

14. Imaging of DNA and Protein by SFM and Combined SFM-TIRF Microscopy
Malgorzata Grosbart, Dejan Ristic, Humberto Sanchez, and Claire Wyman

15. Atomic Force Microscopy of Protein Shells: Virus Capsids and Beyond
Alvaro Ortega-Esteban, N. Martin-Gonzalez, F. Moreno-Madrid, Aida Llauro, Mercedes Hernando-Perez, and Pedro J. de Pablo

16. Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Single Molecule Manipulation and Visualization
Yeonee Seol and Keir C. Neuman

17. Tethered Particle Motion: An Easy Technique for Probing DNA Topology and Transcription Factor Interactions
Daniel T. Tovari, Yan Yan, Laura Finzi, and David Dunlap

18. Single-Molecule Measurements Using Acoustic Force Spectroscopy (AFS)
Douwe Kamsma, and Gijs J.L. Wuite

19. Repurposing a Benchtop Centrifuge for High-Throughput Single-Molecule Force Spectroscopy
Daren Yang, and Wesley P. Wong