BISC219/F13: RNAi Lab 10

From OpenWetWare
Revision as of 18:44, 1 September 2013 by Melissa Beers (talk | contribs)
Jump to: navigation, search

Lab 10: Phenotype Analysis of hsf-1 RNAi worms

Instructors will do 24 hours before lab:
Your instructor or our lab specialist will come in 24 hours before lab and "heat shock" one plate of each of your RNAi samples: wild type treated, wild type control, rrf-3 treated, rrf-3 control, CL2070 treated and CL2070 control. The second plate of each condition serves as your heat shock control. Heat shocking involves moving the worms to a 37°C incubator for 30-45 minutes. The worms will then be placed back at their initial temperature until you use them tomorrow.

To Do in Lab Today
Reverse genetics allows us to start with a known gene sequence and determine the phenotype associated with it. You will determine the effect of RNAi of hsf-1 on several different strains of worms.

We only have one fluorescent compound scope to view the worms and one fluorescent dissecting scope. While some groups are scoring their wild type and rrf-3worms for phenotypes others will be working with the instructor in the microscope room to view and photograph their GFP worms.

  1. Bring your 4 plates of CL2070 worms down to the microscope room.
  2. You will first look at your worms and take some pictures under the dissecting scope. Place one of your non-heat shocked plate on the stage of the dissecting scope and expose the worms to the UV light. Record in your notebook what you see. Are the worms glowing? What part of the worms are glowing?