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Title In vivo mammalian alkaline comet assay
Published [Paris] : OECD, 2014
Online access available from:
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Description 1 online resource (25 pages)
Series OECD guidelines for the testing of chemicals, Section 4: Health effects, 2074-5788 ; test no. 489
OECD guidelines for the testing of chemicals. Section 4, Health effects ; test no. 489
Summary The in vivo alkaline single cell gel electrophoresis assay, also called alkaline Comet Assay is a method measuring DNA strand breaks in eukaryotic cells. Each treated group is composed of a minimum of 5 animals of one sex (or of each sex as appropriate). A positive and a vehicle control group are also used. Administration of the treatment consists of daily doses over duration of 2 days or more, ensuring the test chemical reaches the target tissue which can be the liver, the kidney or other tissues if justified. Tissues of interest are dissected and single cells/nuclei suspensions are prepared and embedded in agarose on slides. Cells/nuclei are treated with lysis buffer to remove cellular and/or nuclear membranes. The nuclear DNA in the agar is then subjected to electrophoresis at high pH. This results in structures resembling comets which by using suitable fluorescent stain, can be observed by fluorescent microscopy. Based on their size DNA fragments migrate away from the head to the tail, and the intensity of the comet tail relative to the total intensity (head plus tail) reflects the amount of DNA breakage
Notes Description based on online resource; title from PDF title page (OECD iLibrary, viewed Dec. 01, 2014)
Subject DNA damage -- Testing.
Gel electrophoresis.
Form Electronic book
Author OECD iLibrary.
Organisation for Economic Co-operation and Development.
ISBN 9264224173 (ebook)
9789264224179 (ebook)