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their up-to-date expertise and experiences, including hundreds of protocols, complete with explanations, and hitherto unpublished troubleshooting hints.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 solutions 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.
विषयसूची
Preface XXXV
List of Contributors XXXVII
Part I RNA Synthesis and Detection 1
1 Enzymatic RNA Synthesis Using Bacteriophage T7 RNA Polymerase 3
Markus Gößringer, Dominik Helmecke, Karen Köhler, Astrid Schön, Leif A. Kirsebom, Albrecht Bindereif, and Roland K. Hartmann
2 Production of RNAs with Homogeneous 5′- and 3′-Ends 29
Mario Mörl and Roland K. Hartmann
3 RNA Ligation 45
Janne J. Turunen, Liudmila V. Pavlova, Martin Hengesbach, Mark Helm, Sabine Müller, Roland K. Hartmann, and Mikko J. Frilander
4 Northern Blot Detection of Small RNAs 89
Benedikt M. Beckmann, Arnold Grünweller, and Roland K. Hartmann
5 Rapid, Non-Denaturing, Large-Scale Purification of In Vitro Transcribed RNA Using Weak Anion-Exchange Chromatography 105
Laura E. Easton, Yoko Shibata, and Peter J. Lukavsky
6 3′-Terminal Attachment of Fluorescent Dyes and Biotin 117
Dagmar K. Willkomm and Roland K. Hartmann
7 Chemical RNA Synthesis, Purification, and Analysis 129
Brian S. Sproat
8 Modified RNAs as Tools in RNA Biochemistry 151
Thomas E. Edwards and Snorri Th. Sigurdsson
Part II Structure Determination 173
9 Direct Determination of RNA Sequence and Modification by Radiolabeling Methods 175
Olaf Gimple and Astrid Schön
10 Probing RNA Structure In Vitro with Enzymes and Chemicals 205
Anne-Catherine Helfer, Cédric Romilly, Clément Chevalier, Efthimia Lioliou, Stefano Marzi, and Pascale Romby
11 Probing RNA Solution Structure by Photocrosslinking: Incorporation of Photoreactive Groups at RNA Termini and Determination of Crosslinked Sites by Primer Extension 231
Michael E. Harris
12 Terbium(III) Footprinting as a Probe of RNA Structure and Metal Binding Sites 255
Dinari A. Harris, Gabrielle C. Todd, and Nils G. Walter
13 Pb2+-Induced Cleavage of RNA 269
Leif A. Kirsebom and Jerzy Ciesiolka
14 Identification and Characterization of Metal Ion Coordination Interactions with RNA by Quantitative Analysis of Thiophilic Metal Ion Rescue of Site-Specific Phosphorothioate Modifications 285
Michael E. Harris
15 Probing RNA Structure and Ligand Binding Sites on RNA by Fenton Cleavage 301
Corina G. Heidrich and Christian Berens
16 Measuring the Stoichiometry of Magnesium Ions Bound to RNA 319
Andrew J. Andrews and Carol A. Fierke
17 Nucleotide Analog Interference Mapping and Suppression (NAIM/NAIS): a Combinatorial Approach to Study RNA Structure, Folding, and Interaction with Proteins 329
Olga Fedorova, Marc Boudvillain, and Christina Waldsich
18 Nucleotide Analog Interference Mapping (NAIM): Application to the RNase P System 369
Simona Cuzic-Feltens and Roland K. Hartmann
19 Identification of Divalent Metal Ion Binding Sites in RNA/DNA-Metabolizing Enzymes by Fe(II)-Mediated Hydroxyl Radical Cleavage 397
Yan-Guo Ren, Niklas Henriksson, and Anders Virtanen
20 RNA Structure and Folding Analyzed Using Small-Angle X-Ray Scattering 407
Nathan J. Baird, Jeremey West, and Tobin R. Sosnick
21 Temperature-Gradient Gel Electrophoresis of RNA 427
Detlev Riesner and Gerhard Steger
22 UV Melting Studies with RNA 445
Philippe Dumas, Eric Ennifar, Francois Disdier, and Philippe Walter
23 RNA Crystallization 481
Jiro Kondo, Claude Sauter, and Benôyt Masquida
24 Studying RNA Using Single Molecule Fluorescence Resonance Energy Transfer 499
Felix Spenkuch, Olwen Domingo, Gerald Hinze, Thomas Basché, and Mark Helm
25 Atomic Force Microscopy Imaging and Force Spectroscopy of RNA 527
Malte Bussiek, Antonie Schöne, and Wolfgang Nellen
Part III RNA Genomics & Bioinformatics, Global Approaches 547
26 Secondary Structure Prediction 549
Gerhard Steger
27 RNA Secondary Structure Analysis Using Abstract Shapes 579
Robert Giegerich and Björn Voß
28 Screening Genome Sequences for known RNA Genes or Motifs 595
Daniel Gautheret
29 Homology Search for Small Structured Non-coding RNAs 619
Manja Marz, Stefanie Wehner, and Peter F. Stadler
30 Predict RNA 2D and 3D Structure over the Internet Using MC-Tools 633
Stephen Leong Koan, Jonathan Roy, Marc Parisien, and Francois Major
31 S2S-Assemble2: a Semi-Automatic Bioinformatics Framework to Study and Model RNA 3D Architectures 667
Fabrice Jossinet and Eric Westhof
32 Molecular Dynamics Simulations of RNA Systems 687
Pascal Auffinger
33 Identification and Characterization of Small Non-coding RNAs in Bacteria 719
Dimitri Podkaminski, Marie Bouvier, and Jörg Vogel
34 The Identification of Bacterial Non-coding RNAs through Complementary Approaches 787
Björn Voß and Wolfgang R. Hess
35 Experimental RNomics, a Global Approach to Identify Non-coding RNAs in Model Organisms, and RNPomics to Analyze the Non-coding RNP Transcriptome 801
Mathieu Rederstorff and Alexander Hüttenhofer
36 Computational Methods for Gene Expression Profiling Using Next-Generation Sequencing (RNA-Seq) 821
John C. Castle
37 Characterization and Prediction of mi RNA Targets 833
Jean Hausser and Mihaela Zavolan
38 Barcoded c DNA Libraries for mi RNA Profiling by Next-Generation Sequencing 861
Markus Hafner, Neil Renwick, John Pena, Aleksandra Mihailovic, and Thomas Tuschl
39 Transcriptome-Wide Identification of Protein Binding Sites on RNA by PAR-CLIP (Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation) 877
Jessica I. Hoell, Markus Hafner, Markus Landthaler, Manuel Ascano, Thalia A. Farazi, Greg Wardle, Jeff Nusbaum, Pavol Cekan, Mohsen Khorshid, Lukas Burger, Mihaela Zavolan, and Thomas Tuschl
40 Global Analysis of Protein–RNA Interactions with Single-Nucleotide Resolution Using i CLIP 899
Julian König, Nicholas J. Mc Glincy, and Jernej Ule
Part IV RNA Function, RNP Analysis, SELEX, RNAi 919
41 Use of RNA Affinity Matrices for the Isolation of RNA Binding Proteins 921
Markus Englert, Bettina Späth, Steffen Schiffer, Sylvia Rösch, Hildburg Beier, and Anita Marchfelder
42 Biotin-Based Affinity Purification of RNA–Protein Complexes 935
Marco Preußner, Silke Schreiner, Inna Grishina, Zsofia Palfi, Jingyi Hui, and Albrecht Bindereif
43 Affinity Purification of Spliceosomal and Small Nuclear Ribonucleoprotein Complexes 957
Julia Dannenberg, Patrizia Fabrizio, Cindy L. Will, and Reinhard Lührmann
44 Study of RNA–Protein Interactions and RNA Structure in Ribonucleoprotein Particles (RNPs) 975
Virginie Marchand, Annie Mougin, Agn´es Méreau, Isabelle Behm-Ansmant, Yuri Motorin, and Christiane Branlant
45 Immunopurification of Endogenous RNAs Associated with RNA Binding Proteins In vivo 1017
Minna-Liisa Änkö and Karla M. Neugebauer
46 Protein–RNA Crosslinking in Native Ribonucleoprotein Particles 1029
Olexandr Dybkov, Henning Urlaub, and Reinhard Lührmann
47 Sedimentation Analysis of Ribonucleoprotein Complexes 1055
Tanja Rösel, Jan Medenbach, Andrey Damianov, Silke Schreiner, and Albrecht Bindereif
48 Identification and Characterization of RNA Binding Proteins through Three-Hybrid Analysis 1067
Felicia Scott and David R. Engelke
49 Experimental Identification of Micro RNA Targets 1087
Michaela Beitzinger and Gunter Meister
50 Aptamer Selection against Biological Macromolecules: Proteins and Carbohydrates 1097
Franziska Peter and C. Stefan Voertler
51 In Vitro Selection against Small Targets 1139
Dirk Eulberg, Christian Maasch, Werner G. Purschke, and Sven Klussmann
52 SELEX Strategies to Identify Antisense and Protein Target Sites in RNA or hn RNP Complexes 1165
Martin Lützelberger, Martin R. Jakobsen, and Jørgen Kjems
53 Genomic SELEX 1185
Jennifer L. Boots, Katarzyna Matylla-Kulinska, Marek Zywicki, Bob Zimmermann, and Renée Schroeder
54 In vivo SELEX Strategies 1207
Thomas A. Cooper
55 Gene Silencing Methods Using Vector-Encoded si RNAs or mi RNAs 1221
Ying Poi Liu and Ben Berkhout
56 Using Chemical Modification to Enhance si RNA Performance 1243
Jesper B. Bramsen, Arnold Grünweller, Roland K. Hartmann, and Jørgen Kjems
Appendix: UV Spectroscopy for the Quantitation of RNA 1279
Index 1283
लेखक के बारे में
Roland K. Hartmann is Professor of Pharmaceutical Chemistry at the Philipps-Universität Marburg (Germany). He studied Biochemistry at the Freie Universität 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 Tübingen 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 m RNA splicing, both in the mammalian system and in trypanosomes.
Astrid Schön received her Ph D from the Institute of Biochemistry at the University of Würzburg, and was a graduate fellow and postdoctoral associate with Dieter Söll 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 Université Louis Pasteur in Strasbourg (France) since 1988. He studied in Liege 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.
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