Spliceosomal Pre-mRNA Splicing: Methods and Protocols [Hardcover]

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.

Comprehensive and practical, Spliceosomal Pre-mRNA Splicing: Methods and Protocols will aid newcomers and seasoned molecular biologists in understanding the fascinating world of alternative splicing with the ultimate goal of paving the way for many new discoveries to come.

The Pre-mRNA Splicing Reaction.- Diversity and Evolution of Spliceosomal Systems.- Mechanisms of Spliceosomal Assembly.- Alternative Pre-mRNA Splicing.- Regulation of Alternative Pre-mRNA Splicing.- Introduction to Co-Transcriptional RNA Splicing.- Chromatin and Splicing.- Preparation of Splicing Competent Nuclear Extracts.- Preparation of Yeast Whole Cell Splicing Extract.- Efficient Splinted Ligation of Synthetic RNA Using RNA Ligase.- In Vitro Assay of Pre-mRNA Splicing in Mammalian Nuclear Extract.- Kinetic Analysis of In Vitro Pre-mRNA Splicing in HeLa Nuclear Extract.- In Vitro Systems for Coupling RNAP II Transcril£‹