Materials:

 

Wild type Drosophila melanogaster larvae

Engrailed-lacZ (En-lacZ)Drosophila melanogaster larvae

Fine forceps

Tungsten needles

Pipettes (10mL, 200 mL)

Pasteur Pipette

Insect Ringer’s solution (Insect saline solution)

Ice bucket

Kimwipe

Vials (for larvae)

Depression slides

Spatula

Eppendorf tubes

Water bath, 37°C

Refrigerator, 4°C

Dissecting microscope

Fixative - 4% PF (paraformaldehyde)

             - 0.1M MOPS (morpholinepropanesulfonic acid), pH7.2

             - 0.01%Tween

               *Must be stored at 4°C. Keep chilled in ice bucket when in use.

X-gal assay buffer 1X -10mM NaPO4 buffer, pH7.2

                                    -1mM MgCl2

                                    -150mM NaCl

X-gal stain solution - X-gal (5-bromo-4-chloro-indolyl-b-D-galactoside)

                              - 4mM K-ferrocyanide (5mM K4Fe(CN)6·3H20)

                              - 4mM K-ferricyanide (5mM K3Fe(CN)6 )

                              - 5mM K2PHO4

                              - 2mM KCl 

                                * Adjusted to pH 7.0. Stored in the dark at 4° to prevent

                                             unwanted reactions within the solution

DMF (dimethylformamide)

Olympus SZ×12 Stereomicroscope w/ DP12 digital camera

 

 

 

 

 

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        Last modified: 4 May 2004

         Laura Carballo & Brian Hwang