BISC 219: Mod 2 Lab 7

Scoring RNAi Worms, Single Worm PCR and Agarose Gel Electrophoresis

3 days before next lab:

1. Come into lab and find your stack of plates.
2. On 2 of the experimental plates add 2 L4 wild type (N2) hermaphrodites
3. On 2 of the experimental plates add 2 L4 rrf-3 hermaphrodites
4. On 1 of the control plates add 2 L4 wild type (N2) hermaphrodites
5. On 1 of the control plates add 2 L4 rrf-3 hermaphrodites
6. Wrap all of your plates in an elastic and stick in your lab day box in the worm incubator set at 23°C
   

In lab today:
Today in lab you will examine your RNAi worms and their progeny.

1. Remove your plates from the incubator.
2. Examine your wild type (N2) and rrf-3 RNAi worms. Do you see any difference between the two strains of worms?
3. Compare your RNAi worms to the control worms - are they the same phenotype? Different?
4. Compare your RNAi worms to worms that have a known mutation in that gene - how do the compare?
5. Score and count approximately 50 worms per plate. What is the proportion of affected to unaffected worms?

Be sure to record all of your results in your lab notebook.

RNAi Schedule of Experiments
RNAi General Information
Media Recipes
Lab 4: Picking your gene to RNAi
Lab 5: Plasmid DNA isolation and transformation
Lab 6: Induction of RNAi plasmid and C. elegans feeding
Lab 8: PCR Reaction Cleanup and Sequencing