6.1: Fly dissection

Submitted by kdorfman on Tue, 11/29/2011 - 19:33

Gene Expression in Drosophila - week 1

Check supplies in the "Drosophila stuff" cupboard under the white board before starting prep.

Fixative

Make fresh each lab day

  • 0.2% glutaraldehyde in Shens

  • 0.05 mL glutaraldehyde in 25 mL Shen's

  • aliquot ~2 mL per group

Stain buffer

ingredient Cf (mM) Stock (M) 5 25 50 mL
NaHPO4 buffer 10 1 0.05 0.25 0.5 mL
K4Fe(II)(CN)6 31 0.005 0.31 1.55 3.1 mL
K3Fe(III)(CN)6 31 0.005 0.31 1.55 3.1 mL
NaCl 150 5 0.15 0.75 1.5 mL
MgCl2 1 1 0.005 0.025 0.05 µL

add a maggot sized pinch per 5 mL

  • aliquot ~2 mL per group

  • can make for both sections at once

Stain

  • pinch of stain in 48 mL stain buffer

  • aliquot 2 mL per group

  • can make for both sections at once

Clearing

1 part stain buffer : 2 parts glycerol

Mounting Medium

ingredient conc Units 100 250 500 1000 mL
gelatin 7 % 7 17.5 35 70 g
glycerol 63 % 63 157.5 315 630 g
  • dissolve gelatine in Shens by boiling
  • Add glycerol
  • Bring to final volume with Shen's
  • Refrigerate