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

Qingzhong Kong | Zendy

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

Author(s) -

Qingzhong Kong

Publication year - 2001

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/29.6.e33

Subject(s) - biology , microbiology and biotechnology , dna , polymerase chain reaction , cloning (programming) , gel electrophoresis , primer dimer , mutagenesis , multiple displacement amplification , ligation , nested polymerase chain reaction , sequencing by ligation , genetics , mutation , dna extraction , base sequence , genomic library , gene , multiplex polymerase chain reaction , computer science , programming language

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.

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