1. GENETICS / Dev. Biol. RSM Embedding, documentation & filing
MODULE 2
The University
of Manchester
Embedding,
documentation
& filing
Life Sciences
Faculty of
Andreas Prokop

2. Background of experiments

3. What is lacZ? The lac operon of E. coli

4. X-gal colour reaction

5. Experiments across the course
Groups
larval genotype
staining
comments
9 - 12, 29
A-lacZ
2hr 37ºC, then o.N. 26ºC
5 groups
B-lacZ
2hr 37ºC
4 groups
, 31, 32
C-lacZ
2hr 26ºC
6 groups
D-lacZ
E-Gal4;UAS-lacZ
5 - 8
F-Gal4;UAS-lacZ
1 - 4
G-lacZ 5

6. β-Gal expressing fly lines
X-lacZ
enhancer
constructs
X-Gal4 UAS-lacZ
enhancer
trap lines
X-lacZ

7. Preparing specimens for analysis

8. Identifying discs upon dissection
head
skeleton
gut
proventricle
on epidermis:
wing, haltere, T3 leg
on CNS:
T1 & T2 leg

9. Embedding of larval preparations
- transfer larval preparations to a drop of glycerol on a slide
- dissect leg and wing discs away and separate on one side of the drop
- place 40 μl droplet of glycerol on a fresh (frosted) slide
- transfer discs with one shank of the forceps (2-5 wing and 2-5 leg discs)
- put on cover slip (see image)
- seal all around the cover slip with nail varnish

10. Filing of slides

11. Documenting specimens

12. Understanding the fate map of leg and wing discs

13. Documentation 18.02.2005 John Smith L3, genotype B X-Gal location: B28
arrow: wing margin
arrow head: wing hinge

14. Your task
Draw as many of the various genetic strains as possible to have material for the assessed task which will be to compose a figure and its legend.

15. Optimising microscope settings