Use of S1 nuclease and formamide in combination for the reassociation studies on GC-rich DNA

Sadhua, Chanchal and Datta, Santanu and Gopinathan, KP (1984) Use of S1 nuclease and formamide in combination for the reassociation studies on GC-rich DNA. In: Analytical Biochemistry, 142 (1). pp. 53-57.

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Abstract

The optimal parameters in the use of nuclease S1 in DNA reassociation kinetics in the presence of formamide have been determined. The conditions are especially suitable for the study of DNA rich in mole percent GC. A 10-fold dilution of the reassociation samples leading to a decrease in both NaCl and formamide concentrations, consequently resulting in a lowering of Tm by only 1.5°C, and the S1 digestion at temperatures identical to the reassociation assay in order to retain the stability of the duplex, are two important aspects of this system. Under these conditions, the kinetics of reassociation followed the theoretically predicted pattern, while the earlier reported methods have shown lower values.

Item Type:	Journal Article
Publication:	Analytical Biochemistry
Publisher:	Elsevier Inc
Additional Information:	Copyright for this article belongs Elsevier Inc.
Keywords:	S1 nuclease; formamide; GC-rich DNA; DNA reassociation; reassociation kinetics
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	02 Feb 2010 12:01
Last Modified:	19 Sep 2010 05:32
URI:	http://eprints.iisc.ac.in/id/eprint/20364

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