Breaksite batch mapping, a rapid method for assay and identification of DNA breaksites in mammalian cells.

Abstract:

:DNA breaks occur during many processes in mammalian cells, including recombination, repair, mutagenesis and apoptosis. Here we report a simple and rapid method for assaying DNA breaks and identifying DNA breaksites. Breaksites are first tagged and amplified by ligation-mediated PCR (LM-PCR), using nested PCR primers to increase the specificity and sensitivity of amplification. Breaksites are then mapped by batch sequencing LM-PCR products. This allows easy identification of multiple breaksites per reaction without tedious fractionation of PCR products by gel electrophoresis or cloning. Breaksite batch mapping requires little starting material and can be used to identify either single- or double-strand breaks.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kong Q,Maizels N

doi

10.1093/nar/29.6.e33

keywords:

subject

Has Abstract

pub_date

2001-03-15 00:00:00

pages

E33

issue

6

eissn

0305-1048

issn

1362-4962

journal_volume

29

pub_type

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