Embedding, documentation

& filing

BIOL22332/20972

GENETICS / Dev.Biol RSM

MODULE 3

The

Uni

vers

ityof

Man

ches

ter

Fac

ulty

of

Life

Sci

ence

s

The lac operon of E. coli

Experimental background

What is lacZ?

Groups larval genotype staining comments

B C A-lacZ2hr 37ºC, then

o.N. 26ºC2 groups

D E B-lacZ 2hr 37ºC 2 groups

F G C-lacZ 2hr 26ºC 2 groups

H I D-lacZ2hr 37ºC, then

o.N. 26ºC2 groups

K L E-Gal4;UAS-lacZ 2hr 37ºC 2 groups, stock a bit weak

M N F-Gal4;UAS-lacZ 2hr 37ºC 3 groups

O P G-lacZ2hr 37ºC, then

o.N. 26ºC2 groups

β-Gal expressing fly lines

X-lacZ

X-lacZ

X-Gal4 UAS-lacZ

enh

ance

r co

nstr

ucts

enh

ance

r tr

ap

lines

on epidermis:

wing, haltere, 3rd leg

on CNS:

1st & 2nd leg

headskeleton

gutproventricle

The transformation of Drosophila imaginal discs into legs and wings

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

Filing of slides

'drawing aid'

18.02.2005John SmithL3, genotype BX-Gallocation: B28

Documentation

arrow: wing marginarrow head: wing hinge

Draw as many of the various genetic strains as possible to have material for the assessed task, i.e. composing a figure and its legend

set to '0'

start small focus of condenser

'A' to 'B'

use little contrast

'C' to 'D'

'B' to 'C'

Groups larval genotype staining comments

B C A-lacZ2hr 37ºC, then

o.N. 26ºC2 groups

D E B-lacZ 2hr 37ºC 2 groups

F G C-lacZ 2hr 26ºC 2 groups

H I D-lacZ2hr 37ºC, then

o.N. 26ºC2 groups

K L E-Gal4;UAS-lacZ 2hr 37ºC 2 groups, stock a bit weak

M N F-Gal4;UAS-lacZ 2hr 37ºC 3 groups

O P G-lacZ2hr 37ºC, then

o.N. 26ºC2 groups