Experimental Procedures
Larval Studies The larvae were acquired from the Honey Bee Research Center located near the University of Guelph. Prior to obtaining the larvae, two 24-well plates were prepared with 300 µL of royal jelly solution. This mixture contained 43.2 mL of Milli-Q water, 14.4 g of lyophilized royal jelly, 1.8 g of glucose, 1.8 g of fructose and 0.6 g of yeast extract. It was stirred in a suitable sized beaker for approximately sixty minutes. Then it was poured into screw-cap veils once it reached a smooth consistency and stored at 4°C until needed. The grafting process consisted of obtaining a frame from one of the hives with larvae at the Honey Bee Research Center. In order to protect the larvae from environmental factors, the frame was wrapped with a towel before the grafting process ensued. With a small grafting tool, the larvae were individually scooped out of the comb on the frame. The larvae were placed into one of the each wells on the twenty-four well plate to float on the royal jelly solution (Figure 9). This was conducted cautiously to ensure that the larva did not flip over since oxygen intake occurs only on one its sides. Once the twenty-four well place …show more content…
A 0.8% agarose gel was made by weighing out 0.24g of agarose and placed into an appropriate sized Erlenmeyer flask. Then 30 mL of 1X TAE buffer was added and the contents were mixed. The mixture was melted in a microwave until it was transparent. Then 3 µL of ethidium bromide was added and the mixture was swirled around. The agarose was poured into the tray and the comb was placed into it after. Once the agarose solidified, the tape was removed carefully to avoid disrupting the comb. It was placed into the electrophoresis chamber with the well positioned near the negative electrode. Afterward, 1X TAE buffer was poured into the chamber until it slightly covered the gel and the comb was removed to expose the