Advanced Experimental Design

1. Advanced Experimental Design Basic Care and Culture of C. elegans

2. Tasks for Today • Pour NGM agar plates for culture of C. elegans • Special kind of growth medium (Nematode Growth Medium) • Pour just like LB agar • Use sterile technique • What should we be sure to do? • Work in groups • Each Student (not group) will need 4-5 plates

3. Basics • Warm up NGM agar • Waterbath, if working • Microwave – CAREFULLY! • Label Petri Dishes with Black Marker • NGM • Date • Your initials • Pour molten NGM into dishes, using sterile technique, let cool

4. Basics - Continued • When plates have cooled/solidified, invert them, wrap in parafilm and put in the incubator in the back of the room that is NOT on.

5. Cultures of OP50 • In order for your C. elegans to eat, we need to feed them some E. coli. • Strain is called OP50 • Grow more slowly than strains used in recombinant DNA technology • We will make broth cultures today and then use them to “Seed” our NGM plates next week so we can culture some C. elegans on those plates

6. What to do: • Obtain a culture tube • Label it OP50, your initials, date • Using sterile technique, transfer 5mls of LB broth into the tube • Using a sterile loop, pick some bacteria from stock culture • Dispose of loop properly (bacterial waste beaker) • Put your labeled tube in incubator in back that is at 37 degrees C.

7. THEN: • Clean up! • Anything that touches bacteria goes in red bin • Wash your hands thoroughly • Write your lab notebook entry