(Ebook PDF) Spliceosomal Pre mRNA Splicing Methods and Protocols 1st Edition by Klemens Hertel ISBN 9781627039802 1627039805 full chapters

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ISBN 10:1627039805

ISBN 13:9781627039802

Author:  Klemens J. Hertel

Providing a guide to classical experimental approaches to decipher splicing mechanisms and experimental strategies that rely on novel multi-disciplinary approaches, Spliceosomal Pre-mRNA Splicing: Methods and Protocols describes the theory of alternative pre-mRNA splicing in seven introductory chapters and then introduces protocols and their theoretical background relevant for a variety of experimental research. These protocol chapters cover basic methods to detect splicing events, analyses of alternative pre-mRNA splicing in vitro and in vivo manipulation of splicing events and high-throughput and bioinformatic analyses of alternative splicing. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls.

Table of Contents:

Theoretical and Mechanistic Studies

1. The Pre-mRNA Splicing Reaction
2. Diversity and Evolution of Spliceosomal Systems
3. Mechanisms of Spliceosomal Assembly
4. Alternative Pre-mRNA Splicing
5. Regulation of Alternative Pre-mRNA Splicing
6. Introduction to Cotranscriptional RNA Splicing
7. Chromatin and Splicing

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Experimental and Methodological Studies

8. Preparation of Splicing Competent Nuclear Extracts
9. Preparation of Yeast Whole Cell Splicing Extract
10. Efficient Splinted Ligation of Synthetic RNA Using RNA Ligase
11. In Vitro Assay of Pre-mRNA Splicing in Mammalian Nuclear Extract
12. Kinetic Analysis of In Vitro Pre-mRNA Splicing in HeLa Nuclear Extract
13. In Vitro Systems for Coupling RNAP II Transcription to Splicing and Polyadenylation
14. Isolation and Accumulation of Spliceosomal Assembly Intermediates
15. Complementation of U4 snRNA in S. cerevisiae Splicing Extracts
16. Expression and Purification of Splicing Proteins from Mammalian Cells

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Advanced Techniques and Computational Methods

17. Single Molecule Approaches for Studying Spliceosome Assembly and Catalysis
18. Quantifying the Ratio of Spliceosome Components Assembled on Pre-mRNA
19. Antisense Methods to Modulate Pre-mRNA Splicing
20. Using Yeast Genetics to Study Splicing Mechanisms
21. Medium Throughput Analysis of Alternative Splicing by Fluorescently Labeled RT-PCR
22. Chromatin Immunoprecipitation Approaches to Determine Co-Transcriptional Nature of Splicing
23. Computational Approaches to Mine Publicly Available Databases
24. Approaches to Link RNA Secondary Structures with Splicing Regulation
25. Methods to Study Splicing from High-Throughput RNA Sequencing Data
26. Global Protein–RNA Interaction Mapping at Single Nucleotide Resolution by iCLIP-Seq
27. Predicting Alternative Splicing

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Klemens Hertel,Spliceosomal,mRNASplicing Method,Protocols