Eppendorf Microinjection of cDNA into Drosophila embryos - Manual (Page 2)

DNA is introduced at 18 °C in Drosophila embryos by
microinjection, into the posterior end of the egg just before
pole cell formation using a manual micromanipulator (Leica
microsystems), Eppendorf programmable microinjector, the
FemtoJet, and ready to use, sterile Femtotip II microinjection
capillaries (see figure 1).
Userguide No 041 | Page 2
‡ Femtotip II microinjection capillaries (Eppendorf AG,
Hamburg, Germany)
‡ Microloader (Eppendorf AG, Hamburg, Germany)
‡ Microscope slide 76x26 mm (Leica Microsystems)
Agar plates with grape juice
‡ Small mesh basket for collecting rinsing and dechorionat-
ing embryos
‡ Double sided tape (tesa SE, Hamburg, Germany)
‡ Embryo collection chambers
‡ Sodium hypochlorite (Sigma-Aldrich, Germany)
‡ Hair dryer
‡ Voltalef oil (VWR, Bruchsal, Germany)
‡ Plasmid purification kit (Qiagen, Hilden, Germany)
Fig. 1: Workstation for Drosophila injection, consisting of a Leica DM IL inverted microscope, a Leica manual micromanipulator and
an Eppendorf programmable microinjector FemtoJet.
Materials and Methods Materials
Instruments
‡ Inverted microscope (Leica DM IL, Leica Microsystems
CMS GmbH, Wetzlar, Germany)
‡ Manual Micromanipulator (Leica microsystems)
Programmable microinjector FemtoJet (Eppendorf AG,
Hamburg, Germany)
‡ Micropipette 0.5-10 µL
‡ Microcentrifuge Eppendorf 5424 (Eppendorf AG,
Hamburg, Germany)