Document Detail


Biosynthetic preparation of 13C/15N-labeled rNTPs for high-resolution NMR studies of RNAs.
MedLine Citation:
PMID:  23065565     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Luigi Martino; Maria R Conte
Related Documents :
1943145 - Prime numbers and the amino acid code: analogy in coding properties.
24556025 - The higher-order structure in the cells nucleus as the structural basis of the post-mit...
15315715 - Periodicity of dna in exons.
9787625 - Conversion problems concerning automated mapping from icd-10 to icd-9.
1943145 - Prime numbers and the amino acid code: analogy in coding properties.
8217155 - Rapid isolation of dna from fossil and museum specimens suitable for pcr.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  941     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2012  
Date Detail:
Created Date:  2012-10-15     Completed Date:  2013-03-18     Revised Date:  2014-02-20    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  227-45     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Acetates / pharmacology
Carbon Isotopes / chemistry
Cell Proliferation
Culture Media / chemistry
Escherichia coli / cytology,  drug effects,  genetics,  metabolism
Formates / pharmacology
Glucose / pharmacology
Glycerol / pharmacology
Hydrolysis
Isotope Labeling
Magnetic Resonance Spectroscopy
Nitrogen Isotopes / chemistry
Phosphorylation
RNA / biosynthesis*,  chemistry*
Ribonucleotides / biosynthesis*,  chemistry*
Substrate Specificity
Transcription, Genetic
Grant Support
ID/Acronym/Agency:
MC_U117533887//Medical Research Council; //Wellcome Trust
Chemical
Reg. No./Substance:
0/Acetates; 0/Carbon Isotopes; 0/Culture Media; 0/Formates; 0/Nitrogen Isotopes; 0/Ribonucleotides; 63231-63-0/RNA; 64-18-6/formic acid; IY9XDZ35W2/Glucose; PDC6A3C0OX/Glycerol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Se-Derivatized RNAs for X-ray Crystallography.
Next Document:  Preparative separation of ribonucleoside monophosphates by ion-pair reverse-phase HPLC.