AzaC treatment of cells? - (Dec/08/2006 )
Hi Everybody,
I am just getting started with analyzing DNA methylation in genomic DNA from humans, however my supervisor would like me to treat cells with Azac. I was wondering does anyone have any references to do this procedure (i.e. what do I dissolve the AzaC with, how long do I need to treat the cells, what concentration should I use..etc). I would greatly appreciate your help, thank you very much in advance
Nael
Hi Nael,
A pubmed search of 5-aza-deoxycytidine or 5-aza-dC will yield a number of papers.
The effect of this drug is very dependent on cell-line some cell lines have cytidine deaminase actrivity and are therefore resistant to the drug. One way to get around this is to use a new drug call zebularine (again this can be found in pubmed).
I make a working stock of 5-axa-dC of 10uM in PBS.
I then performed a titration of this drug on my cells and measured viability with a trypan blue assay to determine the effects on my cells (ie: are they dying or not) before performing any molecular work on them.
Hope this helps.
Nick
Thank you very much methylnick I will look into it .....thanks for the point about some cells being resistant to the drug. I am going to do my research and if I have any further questions, I guess I will be posting them
Regards,
Nael
A pubmed search of 5-aza-deoxycytidine or 5-aza-dC will yield a number of papers.
The effect of this drug is very dependent on cell-line some cell lines have cytidine deaminase actrivity and are therefore resistant to the drug. One way to get around this is to use a new drug call zebularine (again this can be found in pubmed).
I make a working stock of 5-axa-dC of 10uM in PBS.
I then performed a titration of this drug on my cells and measured viability with a trypan blue assay to determine the effects on my cells (ie: are they dying or not) before performing any molecular work on them.
Hope this helps.
Nick