Shroombee! Where have you been all my life!?

And thank you. You helped me confirm my plan and course of action with my project.

Do you feel that contamination is still more likely if the liquid culture vessel lid has a self healing injection port and tyvek over the gas exchange hole?

One love

Thank you guys for kind words.



I prefer agar. Tried LC a couple times, didn't like it. Even when I do a lot of grain I prefer to G2G than to use LC.

Like Shroombee says, LCs have a high contamination risk, they are basically nutrient rocket fuel for any live organism in your container. But high risk can also bring high reward. To do it well you need to nail your sterile technique, and spread your risk by making multiple LCs for each culture. Rubber ports work well, if you can drill/punch clean, tight-fitting lid holes, use quality rubber vacutainer ports, and limit repeat uses of ported lids/rubber ports.

I particularly like doing multiple aliquots of same-culture LCs in vacutainers like in attached pic: good for multi-year storage of prized cultures, with a small footprint. Just take a small drop or needle jab onto agar to revitalize. You can then check for contam, isolate, make mother needle-biopsy drops or wedges for other plates, and go from there.

That will certainly help. But it won't clean up the source of a lot of contamination: the spores themselves. When you germinate on agar, the mycelium typically outruns contaminants. You transfer from the edge of the mycelium growth to new agar plates. That's usually enough to get a clean culture. You can't do that with liquid culture.

LCs are mever a starting place.

This will probably be my last post here on this so as to not detract from the beauty Leo has been so kind to share.

But thank you all for your input. I have yet to encounter any contaminants in the several runs that I've done, so fingers crossed that I have my sterility on point. My goal in using liquid culture is purely to use a single one several times, while moving onto agar in order to maintain certain strains that I've come across and am fond of.

There were some little detail-gems that I really appreciate from you all.

One love

I've only done one LC, I did what most knowledgeable cultivators suggest against, and inoculated sterilized grain soak water with the end of a spore syringe. In this one instance it worked, quicker than spore to grain.

From everything I've read and learned from reading posts of experienced growers, clean agar is king. Any grow should start there. The next time I do a LC (I plan on expanding some myc this way for a monotub PE grow) I will inoculate with clean mycelium from agar. Just a little strand will do, picked up with sterilized tweezers. There's a Tek called Josex Poke where you poke a needle from a sterile water syringe into colonized agar. Large gauge needle like for spore syringes. This takes a tiny piece of myc into the needle. Then squirt water from syringe into your sterilized LC medium to inoculate.

Or heck. Punch out a transfer or cut a wedge and drop it in there.