I want to culture honey bee neurons, the methods include to supplemented it with sugars, I want to know if after added sugars to the medium is recommend to filtered o sterilized again. orhow its the proceed to prepare this medium its better with L15 powder or liquid?
Dear Daniela,
It would be helpful if you included the reference or recipe for the supplemented L15 you are trying to make so that the RG community can be more helpful.
I make a dissection media for neural tissue that requires added sugars. For that, I start with powdered media, add about 90% of the required water, then add the dry form of the required sugar and any other supplements needed. After all the solids are dissolved I pH the solution, then bring the volume to 1 liter (I buy powder to make 1 liter of media). I sterilize the media in the laminar flow hood by filtration through sterile 0.2 micron filters into sterile bottles. I find this is the easiest way to make sure I have the correct pH and concentration of all the ingredients. Also, powdered media may have a longer shelf life than liquid media (I ask the supplier when I order).
Good Luck!
Jill
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