Due to the vital biological importance of RNA and proteins functioning together within a cell, a protocol volume describing experimental procedures to study their interactions should find a home in many laboratories. RNA-Protein Interaction Protocols, Second Edition updates, complements, and expands upon the popular first edition by providing a collection of cutting-edge techniques developed or refined in the past few years along with tried-and-true methods. The expert contributors explore the isolation and characterization of RNA-protein complexes, the analysis and measurement of RNA-protein interaction, and related novel techniques and strategies. Written in the highly successful Methods inMolecular Biology™ series format, the chapters include brief introductions tothe material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tipson troubleshooting and avoiding known pitfalls. Various methods for affinity-purifying cellular or in vitro reconstituted RNPs A collection of methods for studying RNA-protein interaction occurred inside a cell Various methods for assaying protein footprints on RNA Various methods for mapping protein regions involved in RNA interaction New methods including quantifying specific RNAs, designing siRNAs, measuring RNPase activity, etc. INDICE: From the contents Isolation of a Sequence-Specific RNA Binding Protein, Polypyrimidine Tract Binding Protein using RNA Affinity Chromatography.-An Affinity Oligonucleotide Displacement Strategy to Purify RibonucleoproteinComplexes Applied to Human Telomerase.- RNA Affinity Tags for the Rapid Purification and Investigation of RNAs and RNA-Protein Complexes.- Assembly and Glycerol Gradient Isolation of Yeast Spliceosomes Containing Transcribed or Synthetic U6 SnRNA.- Purification of Ribonucleoproteins Using Peptide-Elutable Antibodies and Other Affinity Techniques.- CLIP: Cross-Linking and ImmunoPrecipitation of In Vivo RNA Targets of RNA Binding Proteins.- Quantitative Analysis ofProtein-RNA Interactions by Gel Mobility Shift.- Monitoring Assembly of Ribonucleoprotein Complexes by Isothermal Titration Calorimetry.- Characterization of RNA-Protein Interactions by Phosphorothioate Footprinting and Its Applications to the Ribosome.
- ISBN: 978-1-58829-419-7
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 490
- Fecha Publicación: 01/08/2008
- Nº Volúmenes: 1
- Idioma: Inglés