problem with RNA isolation from leaves - degradated (Oct/28/2006 )

Hi everyone!

Well im starting a project and I have to extract RNA from infected leaves (virus). First, i used two different samples: a plum (mariana type) and other one. But i only get "good" rna from the other one and not from plum. I repeat the same protocol (Trizol) but i cant get any rna, and when i saw it on agarose gel, it always look degradated. I try it with fresh and freezed leaves (-80), but i always look degradated RNA.

Can anybody help me please?

-parafilm-

hi
how do you disrupt your leaves? s it a quick way or a mechanical relative long prep?
the trizol homogenization step can be done at 4° during 15' which minimizes good the RNases

-fred_33-

QUOTE (fred_33 @ Oct 29 2006, 09:55 AM)

hi
how do you disrupt your leaves? s it a quick way or a mechanical relative long prep?
the trizol homogenization step can be done at 4° during 15' which minimizes good the RNases

I do homogenization with liquid nitrogen, and i quickly transfer ( 100 mg aprox) to a eppendorf, then i add 1 ml of trizol reagent and mix (no vortex)

-parafilm-