pH-dependent specific binding and combing of DNA

Recent developments in the rapid sequencing, mapping, and analysis of DNA rely on the specific binding of DNA to specially treated surfaces. We show here that specific binding of DNA via its unmodified extremities can be achieved on a great variety of surfaces by a judicious choice of the pH. On hyd...

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Bibliographic Details
Published in:Biophysical journal 1997-10, Vol.73 (4), p.2064-2070
Main Authors: Allemand, J.F., Bensimon, D., Jullien, L., Bensimon, A., Croquette, V.
Format: Article
Language:English
Subjects:
Adsorption
Binding Sites
Biophysical Phenomena
Biophysics
Chromosome Mapping
DNA - chemistry
Glass
Histidine
Hydrogen-Ion Concentration
In Vitro Techniques
Peptides
Polylysine
Polymethyl Methacrylate
Polystyrenes
Sequence Analysis, DNA
Silanes
Staining and Labeling
Surface Properties
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Summary:Recent developments in the rapid sequencing, mapping, and analysis of DNA rely on the specific binding of DNA to specially treated surfaces. We show here that specific binding of DNA via its unmodified extremities can be achieved on a great variety of surfaces by a judicious choice of the pH. On hydrophobic surfaces the best binding efficiency is reached at a pH of approximately 5.5. At that pH a approximately 40-kbp DNA is 10 times more likely to bind by an extremity than by a midsegment. A model is proposed to account for the differential adsorption of the molecule extremities and midsection as a function of pH. The pH-dependent specific binding can be used to align anchored DNA molecules by a receding meniscus, a process called molecular combing. The resulting properties of the combed molecules will be discussed.
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(97)78236-5