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QUANTITATION OF DNA DAMAGE IN NON‐RADIOACTIVE DNA
Author(s) -
Sutherland Betsy M.,
Ciarrocchi Giovanni,
Ciomei Marina,
Sutherland John C.
Publication year - 1986
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1986.tb04681.x
Subject(s) - dna , dna supercoil , pyrimidine dimer , chemistry , in vitro , dna damage , in vivo , biophysics , molecule , depolymerization , ap site , pyrimidine , nuclear dna , microbiology and biotechnology , biochemistry , biology , dna replication , genetics , mitochondrial dna , polymer chemistry , gene , organic chemistry
Three principal methods have been developed for measuring femtomoles of damage in nanogram quantities of non‐radioactive DNA. Lesions which can be quantified include single and double strand breaks, alkali labile sites including apurinic and apyrimidinic sites, and pyrimidine dimers. The first in vitro method measures the conversion of supercoiled DNA to relaxed or linear molecules, and can detect up to four lesions per molecule. The second in vitro method (supercoil depletion) assesses the fraction of intact linear molecules of homogeneous length, and allows detection of 8 lesions/molecule. The third method, measurement of molecular length distributions of DNAs of heterogeneous length, reveals the extent of DNA damage and repair in vivo or in vitro.