Videos showing how to collect flies with bait; watch in order:
https://youtu.be/iNQu9KCK3wE short video showing a Drosophila (probably hydei) with numerous mites attached (inside bait bottle)
https://youtu.be/YsCIm3EYxd4 short video showing a Drosophila (probably hydei) with numerous mites attached (outside bait bottle)
-How to resuspend new primers:
If they were ordered desalted, add Qiagen EB buffer to achieve a stock concentration of 200uM. For example, if you received 25.1 nmoles, you would add 125.5 uL of EB buffer. The online spec sheet (at IDTDNA) will tell you how much volume you need to add to achieve a final concentration of 100uM. Once you have added the EB buffer, you should to vortex and wait a few minutes for the primer to resuspend. Make sure you label the tube with the date, and the final concentration (e.g. 200uM).
For working stocks, we typically dilute to 10uM. In this case, you should use water to dilute them.
-Preparing fly food (Opuntia Banana recipe for 1 Liter) updated Jan 2020
All ingredients are in the fly kitchen
1L deionized water
10g agar
27.5g yeast flakes
2g Tegosept (same as Benzoic acid, p-hydroxy-, methyl ester; methyl paraben; nipagen; Lexgard M)
47.5g Corn Syrup (we use HEB’s Hill Country Light Syrup)
30g Malt
137.5g bananas (we usually peel them and freeze them)
2.125g Opuntia powder (We are currently using “Starwest Botanicals Nopal Cactus Powder Wildcrafted, 1 Pound”. We used to purchase fresh “nopalito” in cubes at the grocery store, dehydrate them in an incubator @ ~65°C, then grind them in a coffee grinder).
1.5g Propionic Acid (add ~1ml of ethanol to dissolve; you can do it in a petri dish). The propionic acid is in a small tub under the sink. You can use the ethanol from the squirt bottle (usually in fly room).The Propionic acid plus ethanol mix is added once the food is cooling down (see below).
If vials have not been autoclaved, set the autoclave cycle before you begin preparing the food so that they will be ready.
Boil water for ~10 min in covered pot (you can re-use the aluminum foil to cover it)
Weigh all ingredients on Petri dishes except for corn syrup (use small beaker)
Place bananas, opuntia powder and malt in blender. With ladle take a few spoonfulls of the boiled water from the pot and add to blender. Blend and set aside.
Add agar, yeast flakes, corn syrup and Tegosept to pot with boiling water. Boil and mix with the whisk often for ~10 min. (use this time to prepare vials, to start cleaning/washing old vials, and/or to wash utensils)
Carefully pour the pot contents into the blender and blend at low speed (if you do high speed it will spill). Pour blender contents back into pot and boil (at low/med heat) for another ~10 min, mixing often.
Remove the pot from the heat and insert the food thermometer. In the meantime, add water the to the big pot and heat it up. It will be used as a water bath to keep the contents of the small pot warm while you pour vials.
Arrange the vials on clear racks or directly on the bench for pouring. They need to be visible so that you will put the correct amount.
Once the food has reached 145°F (63°C), mix in the Propionic Acid (plus ethanol).
Use ladle to serve food into a small beaker and from the small beaker pour into each vial. Refill the beaker as necessary.
Once all the food from the pot has been poured into vials, use the hot water from the large pot to begin cleaning utensils. Do not leave dirty dishes or benches.
Place the vials in the white plastic rack and cover with clean cheesecloth to prevent contamination by flies and minimize condensation. As soon as the food is settled, vials can be used. Once the vials are cool and their walls as are relatively free of condensation, cover with clean flugs, place the whole tray in a plastic bag (we reuse a white trash bag) and place in the fridge by the window.