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Protocol CitationTina, Yin-Tse Huang 2022. Beetle rearing media. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2lyn48rvx9/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: June 17, 2022
Last Modified: September 13, 2023
Protocol Integer ID: 64771
Abstract
ambrosia beetle rearing

The recipe is for filling c.a. 18⁠—20 falcon tubes (⁠50ml).
First, add dry ingredients
AB
componentamount
cycloheximide0.175 g
Wesson's salt mixture 0.625 g
yeast 4 g
corn starch4 g
casein5 g
agar15 g
sucrose2.5 g
redbay sawdust100 g

Second, add wet ingredients
AB
componentamount
wheat germ oil1.25ml
95% ethanol2.5ml
deionized water290ml

Mix all the ingredients thoroughly and fill in 50ml falcon tubes.
To compress the media, use a clean 15ml falcon tube with a cap on, and press the media until it gets compressed.
Put a double-layered cotton gauze on the mouth of the filled tubes. The cap is pre-drilled with holes that allow aeration and prevent contamination.


Autoclave the filled tube at 121 °C for 30 min.

Put autoclved tubes in an oven at 55°C for two days. (cotton gauze and cap remains on).


Happy beetle rearing!