Document Detail


Electrophoresis of DNA sequencing fragments at elevated temperature in capillaries filled with poly(N-acryloylaminopropanol) gels.
MedLine Citation:
PMID:  9504829     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The performance of poly(N-acryloylaminopropanol) (poly AAP) gel columns, proved to be stable during electrophoresis at elevated temperature, was investigated. The column manufacturing procedure included the preparation of a coating of the inner wall of the fused silica capillary column with linear poly(AAP). Then, a mixture of the AAP monomer, the cross-linker dihydroxyethylenebisacrylamide (DHEBA) and linear poly(AAP) was introduced into the column and in situ polymerized (for preparation of linear gel columns, the addition of DHEBA was omitted). The poly(AAP) columns were first evaluated by electrophoresis of oligonucleotides at room temperature and at 50 degrees C, utilizing 260 nm UV-absorbance detection. In a further evaluation of column performance, samples of T-terminated DNA Sanger fragments from the bacteria Moraxella were separated at 200 V/cm electrical field strength, utilizing a 488 nm argon ion laser and a confocal optical setup for laser-induced fluorescence (LIF) detection. A temperature increase from 25 degrees C to 50 degrees C effectively released a compression of DNA bands. However, for cross-linked poly(AAP) gel columns, the elevated temperature resulted in a considerable reduction of the DNA sequence reading length. When a linear poly(AAP) column was utilized, no detrimental effect of elevated temperature on the separation could be observed.
Authors:
P Lindberg; P G Righetti; C Gelfi; J Roeraade
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Electrophoresis     Volume:  18     ISSN:  0173-0835     ISO Abbreviation:  Electrophoresis     Publication Date:  1997 Dec 
Date Detail:
Created Date:  1998-05-01     Completed Date:  1998-05-01     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  2909-14     Citation Subset:  IM    
Affiliation:
Royal Institute of Technology, Department of Analytical Chemistry, Stockholm, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Acrylic Resins*
Cross-Linking Reagents
DNA Fragmentation*
Drug Stability
Electrophoresis, Capillary
Gels*
Hot Temperature*
Sequence Analysis, DNA*
Chemical
Reg. No./Substance:
0/Acrylic Resins; 0/Cross-Linking Reagents; 0/Gels

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


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