Germline Extraction Staining Protocol

Place about 50ul tetramisole on a 0.1% polylysine slide

Pick about 5-10 worms and place on slide. Push them down so that they stick to the slide.

Using two syringe needles cut open the worms at their vulva (about the middle of the worm) and allow them to push their germlines out

Remove tetramisole with a P200 pipet

Place about 50ul of dye over the worms

Incubate at RT in the dark for 15 minutes

Remove dye

Wash 3x with M9 buffer

Place a drop of glycerol/PPD on side. Cover and seal with clear nail polish.