BISC 219/2009:Gene Mapping: Difference between revisions
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| 9/8 - 9/14 | | 9/8 - 9/14 | ||
|'''Worm Husbandry''': view worm videos; <br> | |'''Worm Husbandry''': view worm videos; <br> | ||
<br> | |||
Examine wild type and mutant worms; practice picking worms;<br> | Examine wild type and mutant worms; practice picking worms;<br> | ||
<br> | |||
Set up first crosses - '''Autosomal vs X-linked test'''<br> | Set up first crosses - '''Autosomal vs X-linked test'''<br> | ||
Mutant Hunt: search "screen" plates for Dpy worms | <br> | ||
'''Mutant Hunt:''' search "screen" plates for Dpy worms | |||
|'''3-4 days later:'''<br> | |'''3-4 days later:'''<br> | ||
Pick 2 "wild type" looking worms from each Autosomal vs X-linked cross to new plates<br> | Pick 2 "wild type" looking worms from each Autosomal vs X-linked cross to new plates<br> | ||
<br> | |||
Examine phenotypes of autosomal vs X-linked F1 generation<br> | Examine phenotypes of autosomal vs X-linked F1 generation<br> | ||
<br> | |||
Examine mutants from your hunt: If Dpy then cross to N2 males | Examine mutants from your hunt: If Dpy then cross to N2 males | ||
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|Score autosomal vs X-linked cross;<br> | |Score autosomal vs X-linked cross;<br> | ||
'''Linkage Test:''' Cross mutant males by 4 tester strains;<br> | '''Linkage Test:''' Cross mutant males by 4 tester strains;<br> | ||
|'''3-4 days later:'''<br> | |'''3-4 days later:'''<br> | ||
'''Linkage Test:''' transfer 2 L4's from each cross to new plates (4 plates total)<br> | '''Linkage Test:''' transfer 2 L4's from each cross to new plates (4 plates total)<br> | ||
|- | |- | ||
!3 | !3 | ||
|9/22 - 9/28 | |9/22 - 9/28 | ||
|'''Linkage Test:'''Score for linkage<br> | |'''Linkage Test:'''Score for linkage<br> | ||
'''Mapping:'''Pick 5 Dumpy worms to individual plates | |||
'''Mapping:'''Pick 5 | |||
|'''3-4 days later:'''<br> | |'''3-4 days later:'''<br> | ||
'''Mapping:''' Pick 3 double mutants to separate plates<br> | '''Mapping:''' Pick 3 double mutants (unc and dyp) to separate plates<br> | ||
'''Complementation:''' Cross your unknown Dpy mutant worms by N2 males | '''Complementation:''' Cross your unknown Dpy mutant worms by N2 males | ||
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'''Complementation:''' Pick males from Complementation cross #1 and mate with known Dpy strains | '''Complementation:''' Pick males from Complementation cross #1 and mate with known Dpy strains | ||
|'''3-4 days later:''' <br> | |'''3-4 days later:''' <br> | ||
'''Mapping:''' Pick | '''Mapping:''' Pick 3-4 heterozygous males to separate plates (8 males total) and mate with homozygous recessive dpy unc worms<br> | ||
'''Complementation:''' Examine cross plates for Dpy males | '''Complementation:''' Examine cross plates for Dpy males | ||
|- | |- |
Latest revision as of 08:26, 28 September 2009
We would like to thank Dr. Phil Hartman of Texas Christian University for developing this series and providing us with the worms for our crosses.
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 and Backcross and Mapping
Lab 4: Mapping Part 2
Lab 5: Score!
Schedule of Experiments
Lab # | Dates | Activity | Outside lab time |
---|---|---|---|
1 | 9/8 - 9/14 | Worm Husbandry: view worm videos;
|
3-4 days later: Pick 2 "wild type" looking worms from each Autosomal vs X-linked cross to new plates |
2 | 9/15 - 9/21 | Score autosomal vs X-linked cross; Linkage Test: Cross mutant males by 4 tester strains; |
3-4 days later: Linkage Test: transfer 2 L4's from each cross to new plates (4 plates total) |
3 | 9/22 - 9/28 | Linkage Test:Score for linkage Mapping:Pick 5 Dumpy worms to individual plates |
3-4 days later: Mapping: Pick 3 double mutants (unc and dyp) to separate plates |
4 | 9/29 - 10/5 | Mapping: cross N2 males with double mutants Complementation: Pick males from Complementation cross #1 and mate with known Dpy strains |
3-4 days later: Mapping: Pick 3-4 heterozygous males to separate plates (8 males total) and mate with homozygous recessive dpy unc worms |
5 | 10/6 - 10-14 | Mapping: Score and determine map distance between unknown Dpy gene and Unc reference mutation |