| Electrophoresis of DNA sequencing fragments at elevated temperature in capillaries filled with poly(N-acryloylaminopropanol) gels. | |
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MedLine Citation:
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PMID: 9504829 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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P Lindberg; P G Righetti; C Gelfi; J Roeraade |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Electrophoresis Volume: 18 ISSN: 0173-0835 ISO Abbreviation: Electrophoresis Publication Date: 1997 Dec |
Date Detail:
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Created Date: 1998-05-01 Completed Date: 1998-05-01 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 8204476 Medline TA: Electrophoresis Country: GERMANY |
Other Details:
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Languages: eng Pagination: 2909-14 Citation Subset: IM |
Affiliation:
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Royal Institute of Technology, Department of Analytical Chemistry, Stockholm, Sweden. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acrylic Resins* Cross-Linking Reagents DNA Fragmentation* Drug Stability Electrophoresis, Capillary Gels* Hot Temperature* Sequence Analysis, DNA* |
| Chemical | |
Reg. No./Substance:
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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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