I have made two modifications in the Ficoll method I originally described in WBG, vol. 3, #2. First, all Ficoll solutions are made 0. 1 M in NaCl to avoid shocking osmoticalIy sensitive worms like unc-29
). Second, after sedimenting worms through 15% w/w Ficoll 400 ( 15', 300 x g), the worms are diluted with an equal volume of 0.1 M NaCl and floated on 35% w/w Ficoll (15', 300 x g) before washing 2x with 0.1 M NaCl. The Ficoll sedimentation removes dead or degenerated worms, cuticles, bacteria. The flotation removes crystalline debris sometimes occurring in worm cultures. We also find that special care to pipette out the last residue of bacterial medium (e.g. 3XD) before resuspending bacteria for worm growth is most conducive to obtaining uncontaminated worms.