treat plant such as rice with 5-azaCyd - how to treat rice with 5-azaCyd (May/18/2005 )
I want to treat my matetials (rice) with 5-azaCyd
because i haven't use than drug before
I don't know how to deal with it
is there anyone who have used that drug before
if you do can you tell me something about that
thanks very much
5-azaCyd is quite highly labile and I store it at -20C some have stored it at -80C.
normally dissolve to a stock concentration of 100mM PBS and store as aliquots for use.
Freeze-Thaw cycles do not help towards the stability of the drug, also it has a short half life and requires replemishment.
An alturnative is to use Zebularine which does not have this problem at all. Do a pubmed on Peter Jones and Victor Marquez as he has some papers with regard to this.
The reason why I chose 5-azaCyd is that I want to reasearch the station of demethylation in rice
futhermore I have bought the drug and I don't know what is the drug you mentionded
so I think 5-azaCyd is the best for me
and I still want to know to use 5-azaCyd
how are you culturing your rice? as seedlings? I have treated cell-lines with great success, 20uM in solution over 3 days works well with CHO cell lines.
it maybe different in rice if you are looking at the whole plant or even seedlings.
I am not clear if 5-azaCyt will be taken up through the roots and have an effect before it is degraded. Best way to find out is to see how they grow with or without the drug, that will give you a clear indication if the drug is working as it has plieotropic effects, you can imagine disrupting the methylation genome wide will be very bad especially in plants with lots of transposable elements.
Hello NICK you are right
because i want to take a cut way
i ask whether there is anyone who have done that before
if there is noone
i have to do it myself although it is a long way
i think it need a long time to find the way how to use that drug and how many i shoud use
i have no experience with working with plants.
Some things to consider:
1. 5-azadC works as a nucleotide anolog so you require actively replicating cells for it to work effectively, so I guess you would look at the roots.
2. you will need to perform a titration I have seen concenctrations as high as 10uM of the drug, I have treated up to 20uM over three days and these have worked (these are animal cell lines!!)
3. because of the short half life, you need to keep adding the drug to the plants.
4. if you are looking at trying to reactivate the drug you may want to also consider combination treatment with HDAC inhibitors such as TSA or valproic acid.