METHOD FOR DETECTING, QUANTIFYING AND MAPPING DAMAGE AND/OR REPAIR OF DNA STRANDS

摘要

Methods and products for detecting in vitro the presence of damage on DNA or the presence of a biological response to damage on DNA at the molecular level. Molecular Combing or other nucleic acid stretching methods are employed together with compounds reacting with DNA, probes binding DNA, or nucleic acid monomers, especially labeled nucleic acid monomers.

主权项

1. A method for detecting the presence or absence of a repaired, damaged, altered or mutated sequence on a nucleic acid comprising:
(a) extracting a one or more nucleic acids from a sample, and optionally rinsing or washing the extracted sample, (b) stretching the at least one nucleic acid in said extracted sample or immobilizing at least one nucleic acid on a substrate in a non-stretched condition, (c) adding a detectable substance to the stretched nucleic acid, which substance positions itself on one or more repaired, damaged, altered, or mutated portions of the stretched nucleic acid by substituting, binding to it, or converting it into a molecular extremity, (d) detecting the detectable substance on the stretched nucleic acid, and optionally (c) detecting the presence of repaired, damaged, altered, or mutated nucleic acid when said substance is detected and detecting the absence of a damaged or repaired nucleic acid sequence when said detectable substance is not detected; or (a) extracting at least one nucleic acid from a sample, and optionally rinsing or washing the extracted sample, optionally rinsing or washing the extracted nucleic acid sample, (b) adding a detectable substance to said nucleic acid for a time and under conditions sufficient for interaction, which substance positions itself on one or more repaired, damaged, altered or mutated portions of the one or more nucleic acids by substituting, binding to it, or converting it into a molecular extremity, optionally rinsing or washing the nucleic acid sample after contacting it with the detectable substance, (c) stretching the at least one nucleic acid in said interacted nucleic acid sample or immobilizing the at least one nucleic acid on a substrate in a non-stretched condition, (d) detecting the detectable substance on the stretched or immobilized nucleic acid, and (e) detecting or diagnosing the presence of repaired, damaged, altered, or mutated nucleic acid when said substance is detected and detecting or diagnosing the absence of a damaged or repaired nucleic acid sequence when said detectable substance is not detected; or (a) treating a sample containing cells prior to extracting nucleic acids from said sample by adding a detectable substance for a time and under conditions sufficient for interaction with nucleic acids, which substance positions itself on one or more repaired, damaged, altered, or mutated portions of the nucleic acid by substituting, binding to it, or converting it into a molecular extremity, (b) extracting a one or more nucleic acids from said sample, and optionally rinsing or washing the extracted nucleic acid sample, (c) stretching or immobilizing on a substrate the at least one nucleic acid in said interacted nucleic acid sample, (d) detecting the detectable substance on the stretched or immobilized nucleic acid, and (e) detecting or diagnosing the presence of repaired, damaged, altered, or mutated nucleic acid when said substance is detected and detecting or diagnosing the absence of a damaged or repaired nucleic acid sequence when said detectable substance is not detected.