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 Resjournal_title
Nucleic acids researchauthors
Kong Q,Maizels Ndoi
10.1093/nar/29.6.e33keywords:
subject
Has Abstractpub_date
2001-03-15 00:00:00pages
E33issue
6eissn
0305-1048issn
1362-4962journal_volume
29pub_type
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