GENETICS / Dev. Biol. RSM Embedding, documentation & filing

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Presentation transcript:

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

Background of experiments

What is lacZ? The lac operon of E. coli

X-gal colour reaction

Experiments across the course Groups larval genotype staining comments 9 - 12, 29 A-lacZ 2hr 37ºC, then o.N. 26ºC 5 groups 17 - 20 B-lacZ 2hr 37ºC 4 groups 25 - 28, 31, 32 C-lacZ 2hr 26ºC 6 groups 13 - 16 D-lacZ 21 - 24 E-Gal4;UAS-lacZ 5 - 8 F-Gal4;UAS-lacZ 1 - 4 G-lacZ 5

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

Preparing specimens for analysis

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

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

Documenting specimens

Understanding the fate map of leg and wing discs

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

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.

Optimising microscope settings