Document Detail


Metal cation mediated-capillary electrophoresis of nucleic acids.
MedLine Citation:
PMID:  20030314     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nucleic acid electrophoresis separation heavily depends upon gel or nongel sieving matrix. Here we propose a metal ion mediated-capillary electrophoresis (MCE-CE) by utilizing the nonspecific interactions of Mg(2+) and Ca(2+) and demonstrate the size, conformation, or sequence based separation and characterization of versatile nucleic acid molecules in free solution. Mg(2+) and Ca(2+) can induce DNA separation at the concentrations as low as 100 and 50 muM, respectively. Noteworthy, the two naturally occurring polymorphisms of one base substitution that may change the secondary folding structure or base stacking can be discriminated by MCM-CE, showing its unique capability of resolving length-identical but conformation-different ssDNA. Benefiting from the achieved separation, we further demonstrate that the folding conformation of oligomers and its change caused by single base substitution can be promptly sensed by online coupled fluorescence polarization. We anticipate that this method will be applicable in length polymorphism analysis, single-strand polymorphism analysis, hybridization analysis, microRNA analysis, and study of protein-nucleic acid interactions and the conformation-function relationship.
Authors:
Tao Li; Dapeng Zhang; Wenru Luo; Meiling Lu; Zhixin Wang; Yuling Song; Hailin Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  82     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-14     Completed Date:  2010-03-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  487-90     Citation Subset:  IM    
Affiliation:
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center For Eco-Environmental Sciences, Chinese Academy of Sciences, Shuangqing Road 18, Beijing 100085, China.
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MeSH Terms
Descriptor/Qualifier:
Cations / chemistry*
DNA, Single-Stranded / analysis*,  chemistry,  isolation & purification
Electrophoresis, Capillary / methods*
Metals / chemistry*
MicroRNAs / analysis,  chemistry,  genetics
Nucleic Acid Conformation
Polymorphism, Single Nucleotide
Chemical
Reg. No./Substance:
0/Cations; 0/DNA, Single-Stranded; 0/Metals; 0/MicroRNAs

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


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