Methods & Specific Aim
- To pick the P0 generation and screen the F2 generation for roller mutants in order to better understand the biological process that controls movement in C. elegans.
- To observe the F3 generation to see if the mutation is hereditary.
- A pick made with a glass Pasture pipet and a piece of 32 gauge platinum wire.
- Nematode Growth Plates "seeded" with OP50 strain of E. coli.
- The OP50 strain of E. coli is a non-pathogenic laboratory strain.
- Ethyl methanesulfonate to alter the DNA of the N2 population of C. elegans by causing aberrant base pairing.
- Pick 5 healthy looking C. elegans from the N2 plate and transfer them each to their own NGM plate. (P0A1-A5)
- Once eggs are laid transfer the 5 N2 C. elegans to 5 more separate NGM plates. (P0B1-B5)
- Once eggs are on the P0B NGM plates the 5 N2 worms are flamed.
- These plates represent the P0 generation.
- The P0 generation will lay eggs, which will be the F1 generation.
- When the F1 generation is big enough to pick, 5-7 F1s will be transfered from each plate to a new plate. (F1A1-A5 & F1B1-B5)
- Once the F1 generation lays eggs, the F1s are flamed.
- The eggs left on the plates are the F2 generation.
- The F2 generation will be screened for the roller mutation.
- 10 mutants are picked and transfered to to mini Petri Plates.
- The F3 generation is observed for mutations.