Simultaneous determination of DNA double strand breaks and DNA fragment size in cultured mammalian cells exposed to hydrogen peroxide/histidine or etoposide with CHEF electrophoresis
- 1 April 1995
- journal article
- research article
- Published by Oxford University Press (OUP) in Carcinogenesis: Integrative Cancer Research
- Vol. 16 (4) , 703-706
- https://doi.org/10.1093/carcin/16.4.703
Abstract
A CHEF (contour clamped, homogenous electric field) assay allowing the measurement of chemically-generated DNA DSBs (double strand breaks), and the simultaneous estimation of the size of the resulting double stranded DNA fragments, in a single gel run, has been developed. This method combines a very high sensitivity for detecting DNA DSBs with a very good resolution over a broad range of megabase—sized DNA. This information can be obtained in a 68 h gel run, a time which is slightly elevated as compared to the CHEF DSB assay (∼20 h), but dramatically reduced as compared to other CHEF protocols utilized for resolving DNA fragments of 0.2–5.7 Mb (5–14 days). Treatment with 5–10 μM etoposide or 50–100 μM hydrogen peroxide/300 μM histidine produced DNA fragments with a mean size of 7.7×105 bp (from ≦ −0.2 Mb) or 4.6×106 bp (from ≧ 5.7–2.2 Mb), respectively.Keywords
This publication has 0 references indexed in Scilit: