1) Preparation (agarose plates and needles)

Preparation:

Agarose Dish

Prepare 1% or 1,5% (w/v) agarose dishes.
Either 1 or 1,5gr agarose in 100ml eggwater (0,6g instant Ocean in 10L water)
Use a blue capped bottle (at least 250ml volume to allow for extra space during boiling)
Boil in a microwave (make sure to remove the cap of the bottle while boiling!!)
Boil for 30-60s and check the turbidity of the agarose solution (repeat until the solution is clear)
Pour app 25ml hot agarose solution per pertridish.
Allow for the agarose to solidify (app. 10-20min)
Use immediately, or store at 4°C in inverted position.
When using dishes that have been stored for a long time make sure to add a small volume of eggwater to the dishes before starting injection (this will prevent the fish from drying out to fast).

Needles:

Make sure that the capillaries have been calibrated on the filament you are using, when you switch either filament or capillary you should determine the ramp value of the capillaries (switch on needle puller, select program, clear, select no, select ramp value, place a capillary and press pull)