C. elegans total RNA extraction.

Materials

• M9
• RNase-free water
• TRIzol
• Ice
• Liquid N₂
• Heat block set to 37°C
• Vortexer
• RNase-free 1.5 mL tubes
• Eliminase or RNase Zap
• Chloroform
• Cold isopropanol
• 75% ethanol
• TE buffer (optional)
• Centrifuge cooled to 4°C

Washing and freeze-cracking worms

1. Grow up the desired number of 10 cm plates. As a general estimate, each 10 cm plate of adult worms yields at least 10 μg of RNA. Scale accordingly across strains.

Example schedule for a bleach-synchronized population

• D1 — pick 3 × 6 cm plates, 5 worms per plate