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PCR-based methods for detecting DNA damage and its repair at the sub-gene and single nucleotide levels in cells

Grimaldi, KA; McGurk, CJ; McHugh, PJ; Hartley, JA; (2002) PCR-based methods for detecting DNA damage and its repair at the sub-gene and single nucleotide levels in cells. MOL BIOTECHNOL , 20 (2) 181 - 196.

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Abstract

Three PCR-based methods are described that allow covalent drug-DNA adducts, and their repair, to be studied at various levels of resolution from gene regions to the individual nucleotide level in single copy,genes. A quantitative PCR (QPCR) method measures the total damage on both DNA strands in a gene region, usually between 300 and 3000 base pairs in length. Strand-specific QPCR incorporates adaptations that allow damage to be measured in the same region as QPCR but in a strand-specific manner. Single-strand ligation PCR allows the detection of adduct formation at the level of single nucleotides, on individual strands, in a single copy gene in mammalian cells. If antibodies to the DNA adducts of interest are available, these can be used to capture and isolate adducted DNA for use in single-strand ligation PCR increasing the sensitivity of the assay.

Type: Article
Title: PCR-based methods for detecting DNA damage and its repair at the sub-gene and single nucleotide levels in cells
Keywords: DNA damage, DNA repair, PCR based method, drug-DNA adduct, single strand ligation PCR, quantitative PCR, MONOCLONAL-ANTIBODY, ANTICANCER AGENTS, SELECTIVE REPAIR, CISPLATIN, STRAND, SEQUENCES, LIGATION, SITE
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Oncology
URI: http://discovery.ucl.ac.uk/id/eprint/108991
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