BISC 219/2009:Gene Mapping

From OpenWetWare
Jump to navigationJump to search
Wellesley College BISC 219 Genetics

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;

Examine wild type and mutant worms; practice picking worms;

Set up first crosses - Autosomal vs X-linked test

Mutant Hunt: search "screen" plates for Dpy worms

3-4 days later:

Pick 2 "wild type" looking worms from each Autosomal vs X-linked cross to new plates

Examine phenotypes of autosomal vs X-linked F1 generation

Examine mutants from your hunt: If Dpy then cross to N2 males

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
Complementation: Cross your unknown Dpy mutant worms by N2 males

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
Complementation: Examine cross plates for Dpy males

5 10/6 - 10-14 Mapping: Score and determine map distance between unknown Dpy gene and Unc reference mutation