Extraction from Beetles

Protocol

Materials

• micropipette
• micropipette tips
• forceps
• scalpel
• razor
• petri dishes
• ethanol in petri dish

Procedure

Beetles: 10 female beetles

1. Remove elytra and clip wings.
2. Put each beetle in 0.5 ml 1X PBS
3. Vortex (or sonicate) and keep/plate solution.
4. In hood, separate beetle abdomen, head, and pronotum
5. Glue abdomen onto a sterile plate, let dry in hood briefly.
6. Meanwhile, put head in 0.5 ml 1X PBS
7. Crush (substitute for extracting mandibular pouches)
8. Open up the abdomen, suck up the contents under metanotum, transfer it in 0.5 ml 1X PBS’
9. Remove gut, put in 0.5 ml 1X PBS
10. and crush.
11. 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.