Schedule and recommended reading :)
NOTE: first lab report due OCT 3. Final Lab report and presentation due DEC. 9-11.

August 26 and 28
Introduction to the nematode C. elegans as a model system to study developmental biology.
Goals:
A. Learn to properly use dissection stereomicroscope (record instructions in your lab note book)
B. Make worm pick and learn to transfer single worms.
C. Start new plate of wild type worms (you will need to do this every lab period).
D. Look at wild type worm plate. Identify adult hermaphrodite, larva stages, eggs (make drawings in you lab note book and label all worms parts you can identify)


September 2 and 4
A. Look at wild type mating plate. Identify males. Make drawings comparing males to hermaphrodites. Label differences. Set up your own mating plate that you will maintain throughout the course. Move 5-10 males to a new plate. Add 3-5 larval hermaphrodites.
B. Describe and quantify wild type worm behaviors: locomotion, defecation cycle, pumping of the pharynx, egg laying, touch sensitivity, etc.
C. Statistics?


September 9 and 11
Anatomical Characterization of wild type worms (Intro to Worm Anatomy)
(See Microscopy Links for tutorials)
Goals:
A. Mount worms on agar pads to view under compound microscope
B. Learn how to use compound microscope, camera, and computer.
C. Take pictures of wild type adult hermaphrodites and males
D. Identify and label all worm parts in your pictures. Use WormAtlas for guidance.


September 16 and 18

Introduction to Differential Interference Microscopy
A. Take pictures of wild type larval stages, eggs and developing embryos.
B. Identify and label all worm parts in your pictures. Use WormAtlas for guidance.
C. Make time lapse movies of cleavage pattern starting with early embryos (1 or 2 cell stage).
D. Time line of gonadal development in hermaphrodite and male.

September 23 and 25

Introduction to Fluorescent microscopy
Flourescent proteins as tools for cell type specific labeling and creating strains with marked cells.
Green Fluorescent Protein History
Other fluorescent labels (Ca sensitive, pH sensitive, voltage sensitive, ATP sensitive)

Goals:
A. Observe worm strain expressing GFP in GABA neurons
B. Pictures of wild type hermaphrodite L1 and L4 GABA nervous system. Make montage and label the identity of all GABA neurons.
C. Design genetic cross to move GFP marker into unc-6 mutant
D. Set up first cross.

September 30 and October 2

FIRST LAB REPORT DUE Oct 3

Goal:
A. DyeI labeling of sensory neurons and (see examples here). This requires coming in the morning before lab and putting your worms in the dye solution.
B. Take pictures of the sensory neurons and label the cell identities.

October 7 and 9

Theory of forward genetic screens; Classical and RNAi
Goals:
A. Identify mutant worms on mutugenized plate. Single (or clone) your suspected mutants. Treat them gently as they may be much more fragile than wild type worms.
B. Anotomical characterization of worm gonad or embryo development. One of these must be included in your lab report.

October 14 and 16

Characterize you mutant worm. Introduction to online resources (Worm Base). (Example from WORM Initiative)
Goals:
A. Characterize differences in behavior and anatomy between wild type and mutant worms. DIC pictures, time lapse, etc.


October 21 and 23

Introduction to Mapping of unknown mutations
A. Mapping to a chromosome. Here is a simple protocol and worksheets (EG1000 and EG1020)for mapping to a chromosome. Begin crosses to mapping strain EG1000 and EG1020. Start by mating you mutant to wild type males (Check out Worm Breeding for Dummies for help with Mapping).

October 28 and 30

Characterize your mutant worms: Fluorescent microscopy
Goals:
A. Set up cross to move GFP chromosome into your mutant.
B. Use of GFP reporter strain to assay cellular phenotypes (see student examples here).
C. Continue crosses to map.
D. Continue behavioral and anatomical comparison of wild type and mutant worms.

November 4 and 6

Theory of confocal microscopy
Use of confocal microscopy to assay cellular phenotypes (see student examples here). Once you have confirmed the GFP chromosome has been moved into you mutant background you can sign up to use the confocal microscope.


November 11 and 13

Gene identification and analysis using WWW tools
Goals:
A. WWW tools to identify gene, location, and possible functions (see Mutant Key)
B. Quantitative analysis of some phenotypic difference between mutant and wild type animals.

November 18 and 20

Continue quantitative analysis of mutant vs wild type phenotypes
.

November 25 (27 is Thanksgiving Holiday)

Present outline of lab report and power point presentatin to TA. Finish gathering pictures and quatitative data for lab report.

December 2 and 4

Run through presentation with instructor or TA

December 9 and 11
Student Power Point Presentations (20 minutes long)

Lab report due in my office (430ASB) by Dec. 123 at 12 noon. Your must also turn in your lab notebook together with you lab report to get full credit.