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• Prep for Worm Lysis
1. Grow worms (chuck them according to the need)
2. Once confluent, collect the worms in one tube & wash them with M9 buffer 2-3 times.
3. Replace the M9 buffer with Lysis buffer and give one wash.
4. Remove as much liquid as possible and freeze the worms in -80*C.
• Worm Lysis
○ 1 bead of protease (purpose of enzyme notes) was added to 10 mL of 1X Native & Non-Native Lysis Buffer. [used in step 2]
1. Thaw the worms on ice
2. Add ~ 0.5 mL - 1 mL of lysis buffer to the tube.
3. Add glass beads as needed (same amt. as the worm pellet.)
4. Perform lysis using bead beater (found in the 4 degree room).
i. 30 sec, 1min incubation, 30 sec.
5. Centrifuge tubes for 15 mins @ 13,000 rpm @ 4*C.
6. Collect the supernatant in new tube and repeat step 5. (This is your worm total lysate)
7. Take ~100 uL for further analysis and store the rest in -80*C.
• SDS Gel:
1. Used the PINKstain Protein Ladder
• Western Blot:
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