Gel electrophoresis of partially denatured DNA. Retardation effect: its analysis and application

The hypothesis about the role of partial denaturatlon in DNA retardation during its electrophoreeis in denaturing gel/1,/2 was tested. We used partially melted DNA molecules in which the size of the melted regions and their location were known. They were obtained through glyoxal treatment of the mel...

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Bibliographic Details
Published in:Nucleic acids research 1982-08, Vol.10 (15), p.4813-4826
Main Authors: Lyamichev, V.I., Panyutin, I.G., Lyubchenko, Yu.L.
Format: Article
Language:English
Subjects:
DNA - isolation & purification
DNA, Bacterial - isolation & purification
DNA, Viral - isolation & purification
Electrophoresis, Polyacrylamide Gel - methods
Indicators and Reagents
Molecular Weight
Nucleic Acid Denaturation
Plasmids
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Summary:The hypothesis about the role of partial denaturatlon in DNA retardation during its electrophoreeis in denaturing gel/1,/2 was tested. We used partially melted DNA molecules in which the size of the melted regions and their location were known. They were obtained through glyoxal treatment of the melted regions by a prooedure allowing the denatured state to be fixed at any point within the melting range. The approach and the availability of the melting maps of DNAs made it possible to investigate DNA molecules differing in length and in the size of the melted regions. The presence of a denatured region at the end of the molecule or inside of it was shown to decrease its electrophoretio mobility, the effect depending on the size of the melted region and on the DNA length. On the basis of the experimental results an explanation is proposed for the cause of retardation in the case of partially denatured DNA.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/10.15.4813