Method of ligation for the detection of abasic sites on a DNA target sequence

Abstract

A method of ligating a ligation product for detecting abasic sites on DNA target sequence is described. The ligation method uses a pyrene probe that ligates with a second probe in the presence of a ligase. A pyrene may comprise 5′-phosphate 1′-pyrene deoxyribonucleotide at the 5′-terminus. The second probe may comprise a terminal 3′-hydroxy, wherein the 3′-hydroxy is ligated with the pyrene probe. The ligase may be T4 DNA Ligase or PBCV-1 as the latter has been found to improve selectivity. The abasic on the DNA target sequence may be naturally occurring or may be a generated abasic that is formed by the addition of an enzyme. Furthermore, selectivity may be increase by the addition of adenosine triphosphate (ATP) or ligating at a temperature above the probe:target sequence thermal disassociation temperature.

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