Document Detail


Characterization of modified and normal deoxyoligonucleotides by MALDI, time-of-flight mass spectrometry.
MedLine Citation:
PMID:  7946298     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A commercially available, matrix-assisted, laser desorption/ionization (MALDI), time-of-flight mass spectrometer has been used for the successful characterization of picomole quantities of modified deoxyoligonucleotides. The procedure was found to be applicable to the analysis of a variety of modified synthetic structures that included oligos with an intact dimethoxytrityl-blocking group, with omega-aminohexyl- and omega-hexyl-modified bases and with alterations to the normal phosphate backbone: 2'-5' and phosphorothioate linkages. Use of an alpha-cyano-4-hydroxycinnamic acid/ammonium formate based matrix gave sharp peaks, sensitivity to 1 pmol, accuracy to 0.06% on average and avoided the necessity for cryogenic technique and/or specialized equipment. Mixtures of normal and modified oligonucleotides worked well without any special precautions as long as the modified oligonucleotides contained a polyanionic backbone. Modified oligonucleotides that contained an amide linkage in place of the usual phosphate ester link required a sinapinic acid- or esculetin-based matrix with no ammonium formate. Crude oligonucleotides could be rapidly analyzed as a quality-control measure using from 10 to 20 pmol of a typical 0.2-mumol scale synthesis.
Authors:
G M Hathaway
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  BioTechniques     Volume:  17     ISSN:  0736-6205     ISO Abbreviation:  BioTechniques     Publication Date:  1994 Jul 
Date Detail:
Created Date:  1994-12-07     Completed Date:  1994-12-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  150-5     Citation Subset:  IM    
Affiliation:
University of California, Riverside.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Lasers
Mass Spectrometry / methods*
Molecular Sequence Data
Oligodeoxyribonucleotides / analysis*
Quality Control
Chemical
Reg. No./Substance:
0/Oligodeoxyribonucleotides

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


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