Beginner with RNA extraction

[K J C 944]

Eduoard,

I tried Chomczynski and Sacchi several times with the same results - low
260-280 ratio.  After making a few small changes in the protocol, I now
get good results (high yield and a ratio >2) EVERY time:

Add phenol (water-saturated) to Solution D at the very beginning.These two
liquids are miscible - you won't see a top and bottom layer.   Add the
tissue sample (ground in liquid N2), vortex and spin.  Transfer  the
supernatant to a new tube, add Chloroform/isoamyl alcohol, vortex, spin. 
At this point you will see two layers.  Keep the top layer and extract
with phenol/chloroform/isoamyl alc. until clear.  Do a final extraction
with just chloroform to remove any phenol left in your sample, then
precip. with isopropanol or ethanol.

Good luck!

Kathy

Kathryn Coyne
College of Marine Studies
University of DE
Lewes, DE  19958