Freezing & Thawing of nematodes

From Wikionchus

Freezing

• wash 4-5 plates (6cm) full of nematodes

(there should be many worms in L2/J1 stage)

with 10 ml freezing solution into a 15ml Falcon tube

• shake gently and pour into four cryo tubes

• place the cryo tubes in a styrofoam box and put the box into a -80°C freezer

• leave for one day at least (max. up to four weeks)

then transfer into liquid nitrogen

Thawing

• take the cryotubes out of the liquid nitrogen
• add 10 ml RPMI 1640 (from GIBCO) when the worms are almost thawn

• fuge gently and discard supernatant

• add 10ml M9
• fuge again
• remove supernatant except for ca. 1ml liquid
• shake gently
• pour worms onto OP50-seeded NGM plates

Links

• Back to Pristionchus pacificus Protocols page