By - WhatWouldJesusBoof
You can most definitely do some transfers from that !!!
1st pic looks good. If that was the first plate you innoculated I would revise your procedure.
Where I failed is I did not keep track of that part…
Are there any other dishes worth saving, or just pretty much let this first one fully colonize and then transfer to a new set of dishes?
I'd transfer some of it now personally.. I also wouldn't open anything you don't want to spread if you don't have to either. I'm pretty inexperienced though so don't take my word for it
About to transfer now! Agar is cooling
Don't put your fingers over the plate.
I did run my gloves down in 70% Isopropyl before doing this. Am I still being risky in doing so?
I meant when you do the transfer.
You need to be very careful to never put anything nonsterile over sterile stuff
It's really ez to have like a tiny piece of your pinker float over open plate and ruin it.
Gotcha. Thank you for the advice!!
What are you using as your mycelium source? Spore syringe, liquid culture syringe, colonized grain?
So it’s absolutely possible to transfer water to agar but success rates are not as good as scraping. Inoculating a grain source first and colonizing it, taking scrapes and rubbing dishes are more effective. The agar looks great. The dishes. The tape. Clearly you’re using sterile tek. It will just come down to water smearing. I honestly would start by getting your liquid culture stash up so you don’t run out of the syringe. From there do grain tests, not agar, then propagate from there. LC isn’t a guarantee and will need to be tested after colonizing, but it looks like you know what you’re doing in terms of cleanliness so I highly recommend starting there.
Could you expound more on the scrapes from grain source? Transferring from grain to agar, I assume? If so, I'd like to know how exactly/what to look for other than rhizomorph. Having trouble googling it.
Sterilize a surgical knife and scrape a small amount of the mycelium from your grain jar. Transfer to agar dish in scraping pattern. A lot of people use dermatology punch tools for circles or surgical knifes for squares or triangles. Really all you need to do is lift a small sample from the rhizo and scrape it into the jelly.
I’ve seen to many people deterred from trading genetics or outright denouncing suppliers for the 3 drop method when really the chances people are getting spores/culture to come out of the needle is limited and who knows what the source of water is in the syringe you’re receiving, and overall standing water is the greatest threat to immature mycelium.
Outside of that, in the comments I see a lot of people talking about finger marks on the lid and while I do agree that handling of the lid should be considered, plate storage should be upside down. Lab journals not only point to the moment of transfer as the leading cause of contamination but that the lid of the plate could have contaminates on it and should be stored upside down. I fill my plates/cups/dishes and stack them while they cool to break down the condensation on the lids. I let them sit over night and inoculate the following day. When I’m done and transfer them to my colonization area, I flip them upside down. Whether it’s straight from the syringe (the water smears but the spore/lc has already stuck to the jelly). Punches as well. But scraping is as good a recommendation I can give to keep unnecessary water off the agar.
Here’s a video to give you an idea and if you look up isolating genetics or cloning, they use a similar method.
Throw 2-9 out, all contam. Pic 1 has that section at the bottom. Transfer that to a new plate ASAP
About to transfer all I can from picture 1 now! Agar is cooling in fresh Petri dishes
Your fingers have been naughty. Srsly now, take care to not hover your hands over open plates. When making transfers try to hold the plates as vertical as you can so that you won't be tempted to hover over the plates.
You have plenty of stuff to work with there. The forst plate looks fine.
Don't open the ones with sporulating mold
If you rly want to you can open the ones with bacteria and transfer what's safe