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Rapid method for detecting base damage in DNA of mammalian cells: assay of U. V. -induced pyrimidine dimers in human cells

Abstract

Simple and rapid techniques are described for the detection of pyrimidine dimers in DNA. Human cells derived from embryonic lung tissue were UV-irradiated and subjected to either an osmotic shock procedure or detergent lysis, then treated with UV-endonuclease from Micrococcus luteus and the DNA partially denatured by treatment with weak alkali. Brief sonication reduced the molecular weight of the DNA, and the single- and double-stranded DNA could then be separated by hydroxylapatite chromatography. Approximately 40% of the expected number of pyrimidine dimers were detected by the enzyme treatment technique. The mean value of numbers of strand breaks per 10/sup 9/ dalton per J/m/sup 2/ was approximately 50% of the expected value. The method has advantages of speed and reproducibility and a large reduction in the quantities of materials used, particularly at the scintillation-counting stage.
Authors:
Bryant, P E; [1]  Jansson, G; Ahnstroem, G
  1. Hammersmith Hospital, London (UK). M.R.C. Cyclotron Unit
Publication Date:
Nov 01, 1978
Product Type:
Journal Article
Reference Number:
AIX-10-448474; EDB-79-096579
Resource Relation:
Journal Name: Int. J. Radiat. Biol.; (United Kingdom); Journal Volume: 34:5
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; DIMERS; CHEMICAL RADIATION EFFECTS; DNA; DOSE-RESPONSE RELATIONSHIPS; EMBRYONIC CELLS; EXPERIMENTAL DATA; GRAPHS; IN VITRO; MOLECULAR WEIGHT; PYRIMIDINES; QUANTITATIVE CHEMICAL ANALYSIS; STRAND BREAKS; TABLES; ULTRAVIOLET RADIATION; ANIMAL CELLS; AZINES; CHEMICAL ANALYSIS; CHEMISTRY; DATA; DATA FORMS; ELECTROMAGNETIC RADIATION; HETEROCYCLIC COMPOUNDS; INFORMATION; NUCLEIC ACIDS; NUMERICAL DATA; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; RADIATION CHEMISTRY; RADIATION EFFECTS; RADIATIONS; 560400* - Other Environmental Pollutant Effects
OSTI ID:
6253256
Country of Origin:
United Kingdom
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: IJRBA
Submitting Site:
INIS
Size:
Pages: 481-488
Announcement Date:

Citation Formats

Bryant, P E, Jansson, G, and Ahnstroem, G. Rapid method for detecting base damage in DNA of mammalian cells: assay of U. V. -induced pyrimidine dimers in human cells. United Kingdom: N. p., 1978. Web. doi:10.1080/09553007814551151.
Bryant, P E, Jansson, G, & Ahnstroem, G. Rapid method for detecting base damage in DNA of mammalian cells: assay of U. V. -induced pyrimidine dimers in human cells. United Kingdom. doi:10.1080/09553007814551151.
Bryant, P E, Jansson, G, and Ahnstroem, G. 1978. "Rapid method for detecting base damage in DNA of mammalian cells: assay of U. V. -induced pyrimidine dimers in human cells." United Kingdom. doi:10.1080/09553007814551151. https://www.osti.gov/servlets/purl/10.1080/09553007814551151.
@misc{etde_6253256,
title = {Rapid method for detecting base damage in DNA of mammalian cells: assay of U. V. -induced pyrimidine dimers in human cells}
author = {Bryant, P E, Jansson, G, and Ahnstroem, G}
abstractNote = {Simple and rapid techniques are described for the detection of pyrimidine dimers in DNA. Human cells derived from embryonic lung tissue were UV-irradiated and subjected to either an osmotic shock procedure or detergent lysis, then treated with UV-endonuclease from Micrococcus luteus and the DNA partially denatured by treatment with weak alkali. Brief sonication reduced the molecular weight of the DNA, and the single- and double-stranded DNA could then be separated by hydroxylapatite chromatography. Approximately 40% of the expected number of pyrimidine dimers were detected by the enzyme treatment technique. The mean value of numbers of strand breaks per 10/sup 9/ dalton per J/m/sup 2/ was approximately 50% of the expected value. The method has advantages of speed and reproducibility and a large reduction in the quantities of materials used, particularly at the scintillation-counting stage.}
doi = {10.1080/09553007814551151}
journal = {Int. J. Radiat. Biol.; (United Kingdom)}
volume = {34:5}
journal type = {AC}
place = {United Kingdom}
year = {1978}
month = {Nov}
}