- 3% Levamisole
- 0.01% PBT (Tween PBS )
- 1.5 ml microtube
- 2 Syringes
- Dissection Plate
- Drawn Pasteur Pippette
- Eppendorf miniSpin Plus
- Pipette 1 mL of 0.01% PBT (Tween PBS) solution into microtube.
- Pick approximately 50 young adult worms into microtube.
- Centrifuge for 60 seconds at 1000 rpm in an Eppendorf miniSpin plus centrifuge
- Aspirate off as much of the liquid as possible with a drawn pasture pipette, taking care not to disturb the worms.
- Wash worms by adding 1 mL of 0.01% PBT to the microtube.
- Centrifuge again for 60 seconds at 1000 rpm and repeat wash.
- Gently resuspend the worms with a micropipetter and transfer worms to glass dissection plate.
- Add an additional 1000 uL of 0.01% PBT solution to glass dissection plate.
- To paralyze the worms, add 2 uL Levamisole (LEV) to the dissection plate. Promptly return LEV to refrigeration. (After 15 minutes, Levamisole changes the morphology of the germline. All dissections must be completed within 15 minutes).
- Place glass dissection plate under microscope and locate worms for dissection.
- Take two syringes and cross tips perpendicularly to worm, with crossing point centered on the base of the head.
- Quickly draw the syringe tips across each other to cleanly cut off head and expose gonad arms (Figure 1).
Cross the needles at the apex of the germ line and drag them across the worm, exposing the germ line.