Lipid extraction from yeast

From LipidomicsWiki

Lipid extraction from yeast (S.cerevisiae)


This method describes how to do total lipid extraction from baker’s yeast. The extracts are analyzed via TLC.


Step 1:
Grow yeast culture to the desired optical density (OD).

Step 2:
Take approximately 75 OD units (for analysis of non labeled lipids) or 7,5 OD units (for analysis of radioactive labeled lipids) and spin down the cells at 4000 rpm for 5 min.

Step 3:
Wash the cells 2x with dH₂O.

Step 4:
Pellet the cells in an 1,5 ml screw cap tube and add 1 ml MeOH/CHCl₃ (2/1) and glass beads. Beat cells in a bead beater for 2x 2min or vortex 5-10 min.

Step 5:
Spin the suspension at maximum speed in a tabletop centrifuge and transfer the supernatant into a new tube.

Step 6:
Add 400 µl 50mM citric acid and 600 µl CHCl₃ to the suspension, mix and spin at maximum speed in a table top centrifuge for 2 min.

Step 7:
Discard upper phase from the tube and dry lower phase in a speed vac or under nitrogen stream. Dissolve the dried lipids in 30-50µl of MeOH/CHCl₃ (2/1) and analyze via TLC.

(for TLC see: Thin layer chromatography (Thiele Lab))