Bligh/Dyer extraction of total lipids:

(Bligh,E.G. and Dyer,W.J. 1959. A rapid method for total lipid extraction and purifcation.
Can.J.Biochem.Physiol. 37:911-917.)
1. Weigh tissue (seed, leaf, etc). Grind dry soybean seeds in ortar and pestle.
!f using leaf or "rabidopsis seeds, add directly to glass tube and sash #ith
glass rod after step $ to grind.
$. "dd tissue to glass tube, add % ls $:1 &e'(:chlorofor. )ash #ith glass
rod.
%. &ix on *utator $+ inutes.
,. "dd 1 l chlorofor and 1.- ls #ater.
.. )ha/e
0. )pin
1. Discard upper layer (a2ueous phase, #ill contain soe polar lipid species li/e
acyl34o"s. Depending on #hat you are after, you ay need to sa5e this layer)
-. 6ransfer lo#er layer (organic phase contains 6"Gs, ebrane lipids and other
neutral lipids) to ne# tube
7. Dry under *$
1+.Dissol5e in $++ ul 1:1 chlorofor: ethanol or 0:1 chlorofor:ethanol.
8!nternal standard3333if using an internal standard, add to step $.