Chemical cleavage reactions of DNA on solid support: application in mutation detectionReport as inadecuate




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BMC Chemical Biology

, 3:1

First Online: 13 May 2003Received: 24 February 2003Accepted: 13 May 2003

Abstract

BackgroundThe conventional solution-phase Chemical Cleavage of Mismatch CCM method is time-consuming, as the protocol requires purification of DNA after each reaction step. This paper describes a new version of CCM to overcome this problem by immobilizing DNA on silica solid supports.

ResultsDNA test samples were loaded on to silica beads and the DNA bound to the solid supports underwent chemical modification reactions with KMnO4 potassium permanganate and hydroxylamine in 3M TEAC tetraethylammonium chloride solution. The resulting modified DNA was then simultaneously cleaved by piperidine and removed from the solid supports to afford DNA fragments without the requirement of DNA purification between reaction steps.

ConclusionsThe new solid-phase version of CCM is a fast, cost-effective and sensitive method for detection of mismatches and mutations.

Keywordssolid phase chemical cleavage reaction mutation detection chemical modification of DNA Electronic supplementary materialThe online version of this article doi:10.1186-1472-6769-3-1 contains supplementary material, which is available to authorized users.

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Author: Chinh T Bui - Andreana Lambrinakos - Jeffrey J Babon - Richard GH Cotton

Source: https://link.springer.com/







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