BISC219/F12: Lab 2: Difference between revisions
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[[BISC219/F12: Lab 7 | Lab 7: Complete Mapping: Score]]<br> | [[BISC219/F12: Lab 7 | Lab 7: Complete Mapping: Score]]<br> | ||
'''Series3:'''<BR> | '''Series3:'''<BR> | ||
[[BISC219/F12: RNAi General Information| Background Information on Project 3: Investigating Gene Regulation Using RNAi]] <br> | |||
[[BISC219/F12: Media Recipes | Media Recipes]]<br> | |||
[[BISC219/F12: RNAi Lab 7 | Lab 7: Identifying a bacterial colony containing our plasmid of interest ]]<br> | |||
[[BISC219/F12: RNAi Lab 8 | Lab 8: Creating the feeding strain of bacteria for RNAi]]<br> | |||
[[BISC219/F12: RNAi Lab 9 | Lab 9: Induction of feeding strain to produce dsRNA and feeding worms]]<br> | |||
[[BISC219/F12: RNAi Lab 10 | Lab 10: Phenotypic analysis of treated vs untreated worms]]<br> | |||
[[BISC219/F12: RNAi Lab 11 | Lab 11: Writing Workshop]]<br> | |||
[[BISC219/F12: RNAi Lab 12 | Lab 12: Writing Conferences]]<br> | |||
</div> | </div> | ||
Revision as of 07:57, 22 August 2012
Lab 2: Finish Project 1- Autosomal or Sex-linked Inheritance?
- 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 hermaphrodite'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.
The main challenge is to correctly differentiate single mutants of each phenotype (Dpy or Unc) from double mutants(Dpy AND Unc). Students tend to undercount single mutants and overcount doubles when they are inexperienced. Check with your instructor if you are unsure about how to score these groups. WT worms may be undercounted because they are harder to "catch". Be careful not to skew your data in this way. Pick what you consider single mutants to a plate and what you consider double mutants to another plate. Ask your instructor to check them before you go to far in the scoring process.
Make sure you copy your data into the appropriate place on the course spreadsheet!
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?).
Assignments
See Assignments page for instructions on Series 1 Results section write up for this project. Due at the beginning of Lab 4. Project 1 Write Up
