| Biosynthetic Preparation of (13)C/ (15)N-Labeled rNTPs for High-Resolution NMR Studies of RNAs. | |
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MedLine Citation:
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PMID: 23065565 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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High-resolution investigations of the structure and dynamics of RNA molecules by nuclear magnetic resonance (NMR) methodologies require the production of (13)C/(15)N-isotopically labeled samples. A common strategy entails the preparation of (13)C/(15)N-enriched ribonucleoside 5'-triphosphates (rNTPs) to be incorporated into RNA oligomers by in vitro transcription. Here, we describe the methods to obtain isotopically labeled rNTP in a uniform or selective fashion from bacterial cultures, using common and versatile E. coli strains. This chapter also covers procedures for extraction and digestion of the total RNA from bacterial cells, purification of the ribonucleoside 5'-monophosphates and their enzymatic phosphorylation to rNTPs. |
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Authors:
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Luigi Martino; Maria R Conte |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 941 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2012 |
Date Detail:
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Created Date: 2012-10-15 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 227-45 Citation Subset: IM |
Affiliation:
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Randall Division of Cell and Molecular Biophysics, King's College London, London, UK. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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