8.2.1. Adults

Procedure:

  1. Place adult bees in filter grinding bag (Neogen™, Bioreba).
  2. Add 0.5 ml grinding buffer (e.g. GITC1) per bee.
  3. Crush the bees.
  4. Transfer 1.5 ml of the supernatant to a 2 ml Eppendorf tube. OR; include a “crude” centrifugation step for bigger volumes.
  5. Centrifuge at 2,000 g for 10 minutes
  6. Transfer 1.5 ml to an Eppendorf tube.
  7. Centrifuge at 20,000 g for 2 minutes.
  8. Discard the supernatant.
  9. Resuspend the pellet in the manufacturer’s lysis buffer (DNeasy® Plant Mini Kit, QIAGEN®).
  10. For manual DNA-extraction: Use DNeasy® Plant Mini Kit (QIAGEN®). Follow the protocol for plant tissue (Mini Protocol). For automated DNA extraction using a QiaCube (QIAGEN®); follow the purification of total DNA from plant tissue standard protocol.
  11. Use the DNA templates directly in a PCR or store in –20 ºC until needed.

1GITC=for 100 ml, add 50 g guanidine thiocyanate, 50 ml nuclease free water, 5.3 ml 1M Tris-Cl (pH 7.6), 5.3 ml 0.2 M EDTA. Stir until completely solved and store at 4 ºC.