BISC 219/2009: Mod 1 Lab 2
Lab 2 Part 1: Scoring Autosomal vs. X-linked
Record in your lab notebook the number of WT, Dpy, Unc and Dpy Unc mutants by scoring the phenotype and removing that animal from the plate (flame the pick to remove the worm). If unlinked, you should see WT's, Dpy’s, Unc’s and Dpy Unc’s in a ratio of 9:3:3:1. If linked, you should see greater than 1/7th Dpy Unc’s among the mutant progeny.
You should now be able to conclude which strain is autosomal and linked, autosomal and unlinked and finally which strain has an x-linked gene and which one it is Dpy or Unc.
Lab 2 Part 2: Linkage Testing and Backcross
Lab 2 Part 3: Calibration of Micropipettes
In this calibration you will measure the absorbance of a blue solution (0.01% BromoPhenolBlue) at various dilutions. The graph of absorbance versus volume of 0.01% BPB should be a straight line.
- Transfer some deionized water from the tap to a 50 ml conical tube. Using your P1000, measure 1 ml of water into each of two 1.5ml plastic cuvettes. They will serve as auto zero blanks for the spectrophotometer.
- Using your P20, measure 10, 15 and 20 microliters of 0.01% BPB solution into the bottom of three more cuvettes. Using your P1000, add water to bring the final volume to 1 ml in each cuvette.
- Using your P200, measure 40, 100 and 200 microliters of 0.01% BPB solution into the bottom of three more cuvettes. Using your P1000, add water to bring the final volumes to 1 ml.
- Using your P1000, measure 201, 300, and 400 microliters of 0.01% BPB solution into the bottom of three cuvettes. Add water to bring the final volume in each cuvette to 1 ml.
- With a gloved hand or with a piece of parafilm over the lip of the cuvette, invert each cuvette several times to thoroughly mix the contents.
- Measure the absorbance of each solution at 600 nm using one of the Hitachi double beam spectrophotometers (requiring two blanks) found in the equipment room, L308.
C. elegans General Information
Tools and Techniques
Lab 1: Welcome to C. elegans and Mutant Hunt
Lab 3: Linkage Test, Backcross and Mapping
Lab 4: Mapping Part 2
Lab 5: Score!
