total RNA analysis - (Apr/02/2007 )
I would like to analyse the total RNA but am very afraid since it so sensitive to nucleases. Can anyone give me advise on how to avoid possible degradation of RNA when analsing it on agarose gel, how to prepare the gel, buffer etc.
I am not that experience in working with RNA. But I will give my 2 cents.
Always prepare your water with DEPC treatment.
Use different sets of pipette or even the tips specialise for RNA work.
And DTT/mercaptho should be able to reduce RNase activity.
Use TBE as your buffer is better for RNA visualisation. I think acrylamide will be the best for gel viewing. Not too sure about the latter gel though.
Hope it helps.