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Dauer Pheromone Prep

Dauer Pheromone Prep

Authors:From Cori Bargmann and Brenda Reinhart 1992
A variation of Riddle's protocol:

Small molecule prep-
1. grow up worms super dense

2. spin worms out

3. filter supe through whatman paper and then through sterile filter

4. boil supe down in a 2 liter flask to about 50 ml

5. spin put insoluble junk, filter and boil to 10 ml

6. pin put junk and boil to dryness-yellowish crust

7. extract it with 10ml ethanol overnight after breaking it up with a spatula into little yellow crusts

8. repeat with several O/N extractions

9. speed vac the etoh to dryness

10. resuspend in 10ml water and freeze

Cleaner prep: but lower yield
1. grow up a mess a worms from 1 liter culture

2. spin out worms and keep them

3. float them through sucrose to get rid of junk

4. put worms in 250ml flask with 50ml s-basal and leave at 20 degrees in shaker o/n

5. spin out worms

6. filter supe and freeze

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Original file name:dauer pheromone prep

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