Titlebook: Spliceosomal Pre-mRNA Splicing; Methods and Protocol Klemens J. Hertel Book 2014 Springer Science+Business Media, LLC 2014 Alternative spli

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In Vitro Assay of Pre-mRNA Splicing in Mammalian Nuclear Extracty to investigate various aspects of splicing and alternative splicing appears to be endless due to the flexibility of this assay. Here, we describe the tools and techniques necessary to carry out an in vitro splicing assay. Through the use of radiolabeled pre-mRNA and crude nuclear extract, spliced

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Inkling

In Vitro Systems for Coupling RNAP II Transcription to Splicing and Polyadenylationy. Recently, in vitro systems were developed for understanding the mechanisms involved in coupling transcription by RNA polymerase II to RNA processing. Here we describe an efficient two-way system for coupling transcription to splicing and a robust three-way system for coupling transcription, splic

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Isolation and Accumulation of Spliceosomal Assembly Intermediatesrategies to arrest spliceosomes at different points of complex formation and provide a detailed protocol developed for isolating intact splicing complexes arrested between the first and second chemical steps of splicing. Briefly, spliceosomes are assembled on a radiolabeled in vitro-transcribed spli

OMIT

Complementation of U4 snRNA in , Splicing Extracts for Biochemical Studies of snRNP Assembly and Funng splicing involves the replacement of endogenous splicing components with their synthetic counterparts. This enables changes in protein or nucleic acid sequence to be tested for functional effects, as well as the introduction of chemical moieties such as cross-linking groups and fluorescent dyes.

commune

Expression and Purification of Splicing Proteins from Mammalian Cellseins. Despite this complexity, the role of individual spliceosomal proteins in the splicing reaction has been successfully investigated using cell-free assays. In many cases, the splicing factor of interest must be expressed and purified in order to study its function in vitro. Posttranslational mod

干旱

Single Molecule Approaches for Studying Spliceosome Assembly and Catalysisey to these experiments is the fabrication of fluorescent molecules (pre-mRNAs and spliceosome components) and passivated glass slides for each experiment. Here we describe how to produce fluorescent RNAs by splinted RNA ligation and fluorescent spliceosome subunits by SNAP-tagging proteins in cell

铁塔等

Book 2014ary 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 b

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