BISC 219/2009: Mod 1 Lab 4: Difference between revisions
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Examine each of the plates initiated last laboratory period. You should see the presence of wild-type hermaphrodites and males. This indicates that the cross was successful. The question is whether there are any Dpy males present. These should be present in roughly equal numbers to the wild-type males if the mutations fail to complement one another. Conversely, if your unknown mutation is allelic to the known Dpy mutation, you should see Dpy males as well. | Examine each of the five plates initiated last laboratory period. You should see the presence of wild-type hermaphrodites and males. This indicates that the cross was successful. The question is whether there are any Dpy males present. These should be present in roughly equal numbers to the wild-type males if the mutations fail to complement one another. Conversely, if your unknown mutation is allelic to the known Dpy mutation, you should see Dpy males as well. | ||
[[BISC 219/2009: Mod 1 C. elegans General Information| ''C. elegans'' General Information]] <br> | [[BISC 219/2009: Mod 1 C. elegans General Information| ''C. elegans'' General Information]] <br> | ||
Revision as of 13:08, 4 September 2009
Lab 4: Mapping and Complementation
Mapping: Cross N2 males X your double mutant (rm). Remember to follow the normal precautions to insure that all progeny are either self progeny (rm mutants) or cis double heterozygous cross progeny (rm/++). Set up duplicate crosses to be sure you get enough worms.
3-4 days after lab:
Pick 2 double heterozygotes to each of two plates for each of your crosses (4 plates total).
Complementation: You will now set up Cross #2: Pick 3-5 L4 and adult males from the plate initiated last week onto five new plates (remember these are heterozygotes for your dpy-u mutation so they are phenotypically wild type but they carry dpy (dpy-u/+). Again, it is essential that only males be transferred onto these plates. Add three L4’s from each of the known dpy strains to your plates.
3-4 days after lab:
Examine each of the five plates initiated last laboratory period. You should see the presence of wild-type hermaphrodites and males. This indicates that the cross was successful. The question is whether there are any Dpy males present. These should be present in roughly equal numbers to the wild-type males if the mutations fail to complement one another. Conversely, if your unknown mutation is allelic to the known Dpy mutation, you should see Dpy males as well.
C. elegans General Information
Tools and Techniques
Lab 1: Welcome to C. elegans and Mutant Hunt
Lab 2: Linkage Test and Backcross
Lab 3: Linkage Test, Backcross and Mapping
Lab 5: Score!
