1 2 3
4
5
6
|
|
|
|
|
|
|
|
|
Materials:
Wild type Drosophila melanogaster larvae Engrailed-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
|
