By Hartmann R.K., et al. (eds.)

This all-in-one compendium of present laboratory thoughts for the examine of RNA brings jointly a variety of novel equipment that have been hitherto in basic terms to be had during the basic literature. The editors - themselves skilled RNA researchers - have amassed specialist authors from the united states and Europe for the final word in daily operating wisdom. To this finish, the entire ideas lined are handled with the person in brain, together with particular useful recommendation and a useful "troubleshooting" part. The huge scope of the e-book permits the side-by-side remedy of specialised tools with easy biochemical recommendations for the isolation, characterization and sensible research of RNA. All pertinent laboratory tools are offered intimately, together with RNA synthesis, amendment and ligation series and constitution decision of RNA Isolation and purification of ribonucleoproteins RNA crystallization unmarried molecule reviews on RNA research of splice variations RNA imaging in vivo Modeling and prediction of RNA constitution RNA microarrays Aptamer libraries and SELEX expertise RNA interference in brief, a consultant to profitable experimentation for molecular biologists, biochemists and biotechnologists and of curiosity to each researcher experimenting with RNA.

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K. Hartmann, A. Bindereif, A. Scho¨n, E. Westhof Copyright 8 2005 WILEY-VCH Verlag GmbH & Co. 1 Enzymatic RNA Synthesis, Ligation and Modification 1 Enzymatic RNA Synthesis using Bacteriophage T7 RNA Polymerase 3 Heike Gruegelsiepe, Astrid Scho¨n, Leif A. Kirsebom and Roland K. 3 1 3 Homogeneous 5 0 and 3 0 Ends, Small RNAs, Functional Groups at the 5 0 End 6 Modified Substrates 7 Transcription Protocols 8 Transcription with Unmodified Nucleotides 8 Transcription with 2 0 -Fluoro-modified Pyrimidine Nucleotides 14 Purification 15 Troubleshooting 17 Low or No Product Yield 17 Side-products and RNA Quality 17 Rapid Preparation of T7 RNA Polymerase 17 Required Material 18 Procedure 18 Cell Growth, Induction and Test for Expression of T7 RNAP 18 Purification of T7 RNAP 19 Notes and Troubleshooting 20 Handbook of RNA Biochemistry.

Stagg and Stephen C. 3 Modeling Dynamic Assembly of the Ribosome with Reduced Representation 556 Conclusion 557 Troubleshooting 557 References 559 Molecular Dynamics Simulations of RNA Systems Pascal Auffinger and Andrea C.

Westhof Copyright 8 2005 WILEY-VCH Verlag GmbH & Co. 3 21 Production of RNAs with Homogeneous 5O and 3O Ends 22 Mario Mo¨rl, Esther Lizano, Dagmar K. Willkomm and Roland K. Hartmann Introduction 22 Description of Approach 23 Cis-cleaving Autocatalytic Ribozyme Cassettes 23 The 5 0 Cassette 23 The 3 0 Cassette 23 Purification of Released RNA Product and Conversion of End Groups 26 Trans-cleaving Ribozymes for the Generation of Homogeneous 3 0 Ends 26 Further Strategies Toward Homogeneous Ends 29 Critical Experimental Steps, Changeable Parameters, Troubleshooting 29 Construction of Cis-cleaving 5 0 and 3 0 Cassettes 29 Dephosphorylation Protocols 33 Protocols for RNase P Cleavage 34 Potential Problems 34 References 35 RNA Ligation using T4 DNA Ligase 36 Mikko J.

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