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by: Hartmann, Roland K.; Bindereif, Albrecht; Schö; n, Astrid; Westhof, Eric

ISBN: 9783527327645 | 3527327649

The second edition of a highly acclaimed handbook and ready reference. Unmatched in its breadth and quality, around 100 specialists from all over the world share their up-to-date expertise and experiences, including hundreds of protocols, complete with explanations, and hitherto unpublished troubleshooting hints. They cover all modern techniques for the handling, analysis and modification of RNAs and their complexes with proteins. Throughout, they bear the practising bench scientist in mind, providing quick and reliable access to a plethora of ... MOREsolutions for practical questions of RNA research, ranging from simple to highly complex. This broad scope allows the treatment of specialized methods side by side with basic biochemical techniques, making the book a real treasure trove for every researcher experimenting with RNA.

VOLUME 1
PART I: RNA Synthesis and Detection
Enzymatic RNA Synthesis Using Bacteriophage T7 RNA polymerase
2 Production of RNAs with Homogeneous 5?- and 3?-Ends
RNA Ligation
Northern Blot Detection of Small RNAs
Rapid, Non-denaturing, Large-scale Purification of in vitro Transcribed RNA Using Weak Anion-exchange Chromatography
3?-Terminal Attachment of Fluorescent Dyes and Biotin
Chemical RNA Synthesis, Purification and Analysis
Modified RNAs as Tools in RNA Biochemistry
PART II: Structure Determination
Direct ... MOREDetermination of RNA Sequence and Modification by Radiolabelling
Probing RNA Structure in vitro with Enzymes and Chemicals
Probing RNA Solution Structure by Photocrosslinking
Terbium(III) Footprinting as a Probe of RNA Structure and Metal-Binding Sites
Pb2+-induced Cleavage of RNA
Identification and Characterization of Metal Ion Coordination Interactions with RNA by Quantitative Analysis of Thiophilic Metal Ion Rescue of Site-specific Phosphorothioate Modifications
Probing RNA Structure and Ligand Binding Sites on RNA by Fenton Cleavage
Measuring the Stoichiometry of Magnesium Ions Bound to RNA
Nucleotide Analog Interference Mapping and Suppression (NAIM/NAIS)
Nucleotide Analog Interference Mapping (NAIM): Application to the RNase P System
Identification of Divalent Metal Ion Binding Sites in RNA/DNA-metabolizing Enzymes by Fe(II)-mediated Hydroxyl Radical Cleavage
RNA Structure and Folding Analyzed Using Small-angle X-ray Scattering
Temperature-Gradient Gelectrophoresis of RNA
UV-melting Studies with RNA
RNA Crystallisation
Atomic Force Microscopy Imaging and Force Spectroscopy of RNA
VOLUME 2
PART III: RNA Genomics & Bioinformatics, Global Approaches
Secondary Structure Prediction
RNA Secondary Structure Analysis Using Abstract Shapes
Screening Genome Sequences for Known RNA Genes or Motifs
Homology Search for Small Structured ncRNAs
Molecular Dynamics of RNA Systems
Identification and Characterization of Small Non-coding RNAs in Bacteria
The Identification of Bacterial non-coding RNAs through Complementary Approaches
Experimental RNomics, A Global Approach to Identify Non-coding RNAs in Model Organisms, and RNPomics to Analyze the Non-coding RNP Transcriptome
Computational Methods for Gene Expression Profiling Using Next-generation Sequencing (RNA-Seq)
Characterization and Prediction of miRNA Targets
Barcoded cDNA Libraries for miRNA Profiling by Next-generation Sequencing
Transcriptome-wide Identification of Protein-binding Sites on RNA by PAR-CLIP (Photoactivatable Ribonucleoside Analog Enhanced Crosslinking and Immunoprecipitation)
PART IV: RNA Function, RNP Analysis, SELEX, RNAi
Global Analysis of Protein-RNA Interactions with Single-nucleotide Resolution Using iCLIP
Use of RNA Affinity Matrices for the Isolation of RNA-binding Proteins
Biotin-based Affinity Purification of RNA-protein Complexes
Affinity Purification of Spliceosomal and Small Nuclear Ribonucleoprotein Complexes
Study of RNA-protein Interactions and RNA Structure in Ribonucleoprotein Particles
Immunopurification of Endogenous RNAs Associated with RNA-binding Proteins in vivo
Protein?RNA Crosslinking in Native Ribonucleoprotein Particles
Sedimentation Analysis of Ribonucleoprotein Complexes
Experimental Identification of microRNA Targets
Aptamer Selection against Biological Macromolecules: Proteins and Carbohydrates
In vitro Selection against Small Targets
SELEX Strategies to Identify Antisense and Protein Target Sites in RNA or hnRNP Complexes
Genomic SELEX
In Vivo SELEX Strategies
Gene Silencing Methods Using Vector-encoded siRNAs or miRNAs
Using Chemical Modification to Enhance siRNA Performance

Roland K. Hartmann is Professor of Pharmaceutical Chemistry at the Philipps-Universit?t Marburg (Germany). He studied Biochemistry at the Freie Universit?t Berlin where in 1988 he received the Ernst Reuter award for outstanding Ph.D. theses. His research interests include natural ribozymes, particularly ribonuclease P, interactions of RNAs with proteins and small ligands, aptamers, antisense and RNA interference.

Albrecht Bindereif is Professor of Biochemistry at the University of Giessen (Germany) since 1999. He studied Biochemistry in T?bingen and at the University of California, Berkeley. His postdoctoral work was done in Michael Green's laboratory at Harvard University. His research interests focus on the mechanism and regulation of mRNA splicing, both in the mammalian system and in trypanosomes.

Astrid Sch?n received her PhD from the Institute of Biochemistry at the University of W?rzburg, and was a graduate fellow and postdoctoral associate with Dieter S?ll at Yale University. She is currently a lecturer at the Institute of Biochemistry, University of Leipzig (Germany). Her research is focused on RNA metabolism and RNA-protein interactions, and the evolution of complex RNA enzymes.

Eric Westhof is Professor of Biophysics at the Universit? Louis Pasteur in Strasbourg (France) since 1988. He studied in Li?ge and Regensburg and was a Fulbright-Hays Research Fellow at the University of Wisconsin with M. Sundaralingam. His research is centered on the relationships between sequence, structure, evolution and functions of RNA molecules.