Lab 2: Finish Series 1- Autosomal or Sex-linked?
- Examine each plate of F2 progeny. If you chose only L4 hermaphrodites, as instructed,you should only see hermaphrodite progeny. If you have a lot of males on your plates, you probably chose young adult worms rather than L4 hermaphrodites's. That's a problem - see your instructor.
- For each cross, you should count and examine a random sample of 100 worms. The mutant worms may be smaller and not move as well as the wild type worms. Look around your plate to get a quick assessment of the population.
- Record in your lab notebook the number of WT, Dpy, Unc, or Dpy Unc mutants by examining the phenotype as you remove each animal from the plate. (Be sure to flame the pick after removing each worm.) If the genes responsible for the mutations are unlinked, you should see WT's (+/+;+/+), Dpy’s (dpy/dpy;+/+), Unc’s(+/+;unc/unc) and Dpy Unc’s (d/d;u/u) in a ratio of 9:3:3:1. If linked, you should see a greater proportion than expected of Dpy Unc’s (d u/d u) double mutants vs Dpy or Unc single mutants among the mutant hermaphrodite progeny.
Make sure you copy your data into the appropriate place on the course spreadsheet on the middle computer in the back of the lab!!!1
You should now be able to conclude which strain has mutations that are autosomal and linked, which strain has mutations that are both autosomal and unlinked, and which strain has an autosomal mutation and an x-linked mutation responsible for either the Dpy or Unc phenotype (which one?).
**See Assignments page for instructions on Series 1 Results section write up for this project. Due at the beginning of Lab 4.
Links to Labs& Project Info
Lab 1: Worm Boot Camp & Sex-Linked or Autosomal Start
Lab 2: Sex-Linked or Autosomal Finale
Background: Classical Forward Genetics and Gene Mapping
Lab 2: Mutant Hunt
Lab 3: Linkage Test Part 1
Lab 4: Linkage Test Part 2, Mapping and Complementation
Lab 5: Finish Complementation; Mapping Con't
Lab 6: DNA sequence analysis; Mapping Con't
Lab 7: Complete Mapping: Score
Schedule of Reverse Genetics Project
RNAi General Information
Lab 5: Picking your gene to RNAi
Lab 6: Cloning your gene of interest
Lab 7: Picking your transformant
Lab 8: Plasmid purification and transformation
Lab 9: Induction of bacteria for RNAi
Lab 10: Scoring your worms and RNA purification
Lab 11: RT PCR reactions