10.3.1. RNAi in adult honey bees via feeding

  1. Take newly emerged bees (1-2 d old) from one colony from one brood frame.
  2. Set up at least two controls:
    1. Bees fed with 50% sugar water alone.
    2. Bees fed with scrambled siRNA (siRNA with exactly the same nucleotides as the target siRNA but in an altered order lacking any similarity with other known bee genes).
  3. Take a mixture of two siRNAs specific for the target gene.
    Note: in previous experiments a mixture of two siRNAs was more effective than single siRNAs (Jarosch et al., 2011).
  4. Put 35-40 newly emerged bees in wooden cages (see the BEEBOOK paper on maintaining adult honey bees in vitro under laboratory conditions (Williams et al., 2013)) supplied with a small comb and pollen ad libitum.
  5. Put cages in temperature controlled incubators (see the BEEBOOK paper on maintaining adult honey bees in vitro under laboratory conditions (Williams et al., 2013)) and feed with 1.5 ml 50% sugar water containing approximately 1µg siRNA per insect every 24 hours.
  6. Dependent on the actual experiments bees can be held for several weeks.
  7. Once the experiment is finished, bees should be shock-frozen in liquid nitrogen in order to maintain the RNA status.