Cleaning worms by floating on sucrose

 

Cleaning worms by floating on sucrose

Have cold 70% sucrose in water.

Wash worms off plates, put on ice.

 

Typically we use 2 - 10 10-cm plates of crowded or starved worms
For this many worms, use a 15ml orange-cap centrifuge tube
Wash off with M9, S-basal, or water
Be sure to grow worms on agarose plates when making DNA to be cut

Wash worms in water.

 

This is optional, it makes them more buoyant
For a 15ml tube, spin in clinical centrifuge at #3 or 4

Mix and equal volume of worms and sucrose.

 

30 - 35% sucrose final, 7 - 10 ml total
Keep cold, worms will thrash

Spin 5 min.

for a 15ml tube, spin in clinical centrifuge at #6 or 7

Draw off floating worms with a pasteur pipette.

Wash worms free of any sucrose.
If worms are to be kept alive, be very careful to use aseptic technique

Bacteria, dead worms and other dense things will be on the bottom