10.3.2. RNAi in honey bee larvae via feeding

  1. Take a comb with second instar larvae out of the colony.
  2. Transfer it to the lab.
    The whole treatment is conducted at room temperature.
  3. Draw a map of the different treatment groups on the very same comb for future identification of the treated individuals.
  4. Apply 1 µl of sugar solution containing the respective amount of dsRNA directly into the cells.
     Deposit it at the bottom of the worker brood cell that contains a drop of food. Avoid touching the larvae. Successful experiments used dsRNA concentrations between 0.5 µg (Nunes and Simões, 2009) and up to 1.26 µg (Aronstein et al., 2006).
    In addition to the first dsRNA feeding you may feed another µg of your dsRNA after 12 hours. This feeding cycle can be repeated for several days (Liu et al., 2010) until the life stage of interest is reached.
  5. Place the comb back to its host colony two hours after treatment and take samples at the life stage you are interested in.