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This updated book reflects improvements in a variety of techniques used to study the aptamer field. Beginning with a section on selection procedures, the volume continues with methods to characterize aptamers' interaction and structural properties by biophysical approaches, as well as a variety of applications that have been adapted to the aptamer compound class. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Nucleic Acid Aptamers: Selection, Characterization, and Application, Second Edition serves as an ideal guide for researchers aiming to further our understanding of aptamer biology and more.
List of contents
Automated ssDNA SELEX Using Histidine Tagged Target Proteins.- Discovery of Aptamers against Cell Surface Markers Using Ligand-Guided Selection.- Implementation of Emulsion PCR for Amplification of Click-Modified DNA during SELEX.- Preparation of Chemically Modified DNA Library for SELEX via Incorporation of CLB-dUTP in Primer Extension Reaction.- RNA Capture-SELEX on Streptavidin Magnetic Beads.- The Bioinformatics of Aptamers: HT-SELEX Analysis with AptaSUITE.- Improvement of Aptamers by High-Throughput Sequencing of Doped-SELEX.- Analysis of Aptamer-Small Molecule Binding Interactions Using Isothermal Titration Calorimetry.- Determination of Aptamer Structure Using Circular Dichroism Spectroscopy.- Immobilization-Free Determination of Dissociation Constants Independent of Ligand Size Using MicroScale Thermophoresis.- Label-Free Determination of the Kinetic Parameters of Protein-Aptamer Interaction by Surface Plasmon Resonance.- Probing of Fluorogenic RNA Aptamers via SupramolecularFörster Resonance Energy Transfer with Universal Fluorescent Nucleobase Analog.- Slow-Off-Rate-Modified Aptamer Labeling for Fluorescence Microscopy and DNA-PAINT.- Aptamer-Field-Effect Transistors for Small-Molecule Sensing in Complex Environments.- Aptasensor for Impedimetric Detection of Lysozyme.- Biophysical Analysis of Small Molecule Binding to Viral RNA Structures.- Designing and Expressing Circular RNA Aptamers to Regulate Mammalian Cell Biology.- Enzyme-Linked OligoNucleotide Assay (ELONA).- High-Throughput Development and Optimization of RNA-Based Fluorogenic Biosensors of Small Molecules Using Droplet-Based Microfluidics.- Robotic APTamer-Enabled Electrochemical Reader (RAPTER) System for Automated Aptamer-Mediated Electrochemical Analysis.
