Wet Plate Protocol for the production of "dauer cultures"

From Wikionchus

To get dauers on wet NGM agar plates - Based on the protocol from Andreas Weller

Protocol

1. Take 1 ml pellet of clean mix-stage worms or bleached/syncronized J2 larvae.
2. Add 50ml of OP50.
3. Take fresh 10cm NGM plates (cold) from 4C room and put them in hood.
4. Seed 1ml of worms+OP50 mixture on the cold 10cm NGM plates in the hood.
5. Put at 25C for faster growth.
6. Monitor plates every 2 days and add 1ml distilled water if the agar surface looks dry.
7. You may add 1 ml OP50 at day 5 to get higher number of worms.
8. The plates should have a large fraction of dauers around days 10 to 12.
9. Do Dauer purification by Sucrose Floatation and Ficoll precipitation or using modified dauer purification protocol.

Links

• Back to Dauer cultures and Purification page
• Back to Pristionchus pacificus Protocols page