- micropipette tips
- petri dishes
- ethanol in petri dish
Beetles: 10 female beetles
- Remove elytra and clip wings.
- Put each beetle in 0.5 ml 1X PBS
- Vortex (or sonicate) and keep/plate solution.
- In hood, separate beetle abdomen, head, and pronotum
- Glue abdomen onto a sterile plate, let dry in hood briefly.
- Meanwhile, put head in 0.5 ml 1X PBS
- Crush (substitute for extracting mandibular pouches)
- Open up the abdomen, suck up the contents under metanotum, transfer it in 0.5 ml 1X PBS’
- Remove gut, put in 0.5 ml 1X PBS
- and crush.
- remove contents from mycangium and transfer in 0.5 ml 1X PBS.
Sonicator test: use six beetles, sonicate two (separately) for 10, 30, 60 seconds. proceed as above. – NO DIFFERENCE, BEETLE STILL LIVING!
Bead beater – fill tubes with 500 μl of PBS. More liquid prevents movement of beads.