Comet assay
Jieru Egeria Lin
Sunday, 28 October 2007 06:18 UTC
I am trying to detect double strand DNA damage using comet assay. Anyone have suggestions about DS DNA damage detection? Is comet assay a good index for DS DNA damage? Also, what should I pay attention to when doing this assay? Thank you very much!
-
Replies
-
Hi Jieru Lin
The comet assay is an excellent assay to use to detect double strand breaks as well as single strand breaks and DNA crosslinks. The neutral comet assay is the one most frequently used for measuring DSB’s and lots of detail can be found in Nature protocols June 2006 DOI 0.1038/nprot.2006.5 This is an excellent article by P. L. Olive and J. P. Banath. Peggy Olive’s group has many years experience of doing the neutral comet assay and a pub med of her papers should give you all the information you need. Another method of estimating DSB is to stain for gama H2AX using IHC or flow cytometry. A nice paper copmparing the two has just appeared on line in Nucleic Acids Research, 2007, Vol. 35, No. 5 e36. A number of articles have been published comparing these tow methods.Hope this helps.
Tim
-
