Hi,
I would like to quantify global chromatin compaction using MNAse treatment followed by Comet assay. However, I don't know if Comet assay would sensitive enough to detect effect of drugs on chromatin compaction.
Has anyone tried it after treatment with chromatin drugs, such as TSA, DNA MethylTransferase inhibitors....
Thank you for your help.
François
Salut François, Je suis pratiquement sûre que l'essai comet n'est pas assez sensible. Ce qu'on faisait après la MNase c'est un gel ADN pour visualiser les coupures entre nucleosomes permettant de quantifier la compaction.
A+ et bonne continuation.
Béa.
Hi François, I wonder whether you have tried the assay and did it work. From my experience, MNAse is relatively insensitive to chromatin compaction. I would guess that DNAse I is better suited. I like your idea and I think that the combination of DNase I with comet assay should work. One would need to titrate DNAse and have both cell types (with relatively condensed vs relaxed chromatin) on the same slide (internal control for each DNase concentration).
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