Has anyone tried miRNA isolation using the flash PAGE fractionator from Ambion? - (Jul/12/2006 )
Has anyone tried the flash PAGE fractionator from Ambion? Its seems quick and easy but is it really efficient?
According to the specs it looks like you only get 10ng miRNA from 100ug of total RNA. Is this what I should expect regardless of the method I use to isolate miRNA?
I just got my miRACLE miRNA Isolation Kit (Stratagene) today. I haven't used it yet, but this should be working very well.
I don't know the exact procedure of yout kit, but usually RNA is fractionated by differences in percentage of ethanol and binding characterisitcs to silica membranes in the presence of chaotropic salts.
I can let you know how my kit wors in about a few days, if you'd like to.
the flash page system can be tedious, or so I've heard. I've used the Ambion mirVana kit and it works well for small RNA purification for many different tissues and cell lines. Much faster as well!
Ive got the flash page, i tried using it with my small RNA fraction to purfy it further & lost alot through the procedure, however ive since found out (like u correctly said) i should be using total RNA to put through the flashpage. However i think the MiRVana kit is efficient enough in itself with the small RNA isolation protocol not to bother with the flashpage! If u do use it & get high yields of small RNA let me know.