Introduction to Optical Tweezers: Background, System Designs, and Commercial Solutions -- RNA Unzipping and Force Measurements with a Dual Optical Trap -- Protein Tethering for Folding Studies -- Combining Structure-Function and Single-Molecule Studies on Cytoplasmic Dynein -- A Brief Introduction to Single-Molecule Fluorescence Methods -- Fluorescent Labeling of Proteins -- Single-Molecule Imaging of Escherichia coli Trans-Membrane Proteins -- Single-Molecule Fluorescence Microscopy in Living Caenorhabditis elegans -- Purification and Application of a Small Actin Probe for Single-Molecule Localization Microscopy -- Fluorescence Microscopy of Nanochannel-Confined DNA -- Use of Single Molecule Fluorescence Polarization Microscopy to Study Protein Conformation and Dynamics of Kinesin--Microtubule Complexes -- Single Molecule FRET Analysis of DNA Binding Proteins -- Atomic Force Microscopy: An Introduction -- Imaging of DNA and Protein by SFM and Combined SFM-TIRF Microscopy -- Atomic Force Microscopy of Protein Shells: Virus Capsids and Beyond -- Combined Magnetic Tweezers and Micro-Mirror Total Internal Reflection Fluorescence Microscope for Single Molecule Manipulation and Visualization -- Tethered Particle Motion: An Easy Technique for Probing DNA Topology and Transcription Factor Interactions -- Single-Molecule Measurements Using Acoustic Force Spectroscopy (AFS) -- Repurposing a Benchtop Centrifuge for High-Throughput Single-Molecule Force Spectroscopy. .

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. .