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sasquanch
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Very new to this - need help identifying if this is contam?
#28688439 - 03/06/24 08:48 AM (3 months, 19 days ago) |
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I just started this fun and exciting journey, and I'm already obsessed.
As kind of an experiment, I wanted to see if I could add liquid culture to agar cups to start the process and control some of the contamination / increase number of healthy mycelium to transfer to grain.
I made the agar and used ketchup cups. None of the agar cups that were extra have started growing any contam.
I DO think I put in too much liquid culture into my test cups, so maybe that's where I screwed up most. This particular cup is has a lot of growth, but I can't tell if it's contamination or if maybe the food coloring I put in the agar was too much, if the extra liquid from the culture is doing weird stuff, or what's going on. It looks to me like the white stuff might be healthy, but I see a sort of blue/green center on those thicker growths.
Any thoughts / ideas?


Thank you!
-------------------- sovereign.church
Edited by sasquanch (03/06/24 08:57 AM)
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Very new to this - need help identifying if this is contam? [Re: sasquanch]
#28688483 - 03/06/24 09:25 AM (3 months, 19 days ago) |
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The thicker growth definitely looks like mold to me.
Putting liquid culture to agar is a great way to test your culture and check how clean it is, as you can see. But if you have a healthy, clean liquid culture, you can inoculate grains with the liquid culture directly. You do not need to use agar for that part.
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  Important Links. Be kind to others.
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Shroomerquest



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Re: Very new to this - need help identifying if this is contam? [Re: sasquanch]
#28688501 - 03/06/24 09:45 AM (3 months, 19 days ago) |
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Did you use a SAB to do the agar work?
Problem here is the liquid culture could be clean but your sterile technique when doing agar work needs honing in.
The thick growths look like mold also, and that could be down to technique or how they’re stored.
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 Relax…yOu’re triPping
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sasquanch
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Re: Very new to this - need help identifying if this is contam? [Re: Shroomerquest]
#28688517 - 03/06/24 10:19 AM (3 months, 19 days ago) |
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Yep, used a SAB, but only wiped down the cups with 70% iso since they couldn't be PC. My cups had a lot of condensation on the sides, but I stacked them up while warm after reading that can help with some of that issue. Left them out for 2 days at room temp to see if I could catch any early contam but nothing showed up.
I'm positive I need some practice with the technique though. Seems weird to me to have to pull my arm in and out of the box to flame sterilize tools frequently. I'm gonna build a flow hood/box soon so I don't have to work in such a small space. It's also probably part of where I'm working (In the kitchen)
-------------------- sovereign.church
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Smellyhobbit
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Re: Very new to this - need help identifying if this is contam? [Re: sasquanch]
#28688535 - 03/06/24 10:44 AM (3 months, 19 days ago) |
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When you say you wiped down the cups, what do you mean?
-------------------- A Love Letter to New Growers A Guide for New Growers Need Spores? - Sablabs.org Just because your tub contamed, doesn’t mean your attitude has to contam as well.

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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Very new to this - need help identifying if this is contam? [Re: Smellyhobbit]
#28688544 - 03/06/24 10:52 AM (3 months, 19 days ago) |
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It sounds like you are saying you took a plastic cup, wiped it out with 70% isopropyl, and then poured hot agar into it.
If that's what happened, you need to be using cups that are truly sterile. Either buy sleeves of pre-sterilized petri dishes on Amazon, or buy some polypropylene cups and switch to a proper no-pour TEK with a pressure cooker.
Do you own a pressure cooker? I'm guessing you used one for sterilizing your agar before pouring it.
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Camera93
We got dicks like Jesus



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Re: Very new to this - need help identifying if this is contam? [Re: Smellyhobbit]
#28688549 - 03/06/24 10:54 AM (3 months, 19 days ago) |
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I'm not sure if you've seen these or not. There are a few videos of members working in a SAB for reference.
Bods video collection
I agree with above assessments, it looks like mold to me too
Crackatoa has a write up on using those ketchup cups : crack plates
-------------------- All I need are some tasty waves, a cool buzz, and I’m fine. Whatever you decide won’t really impact our survival Close your eyes, and do the best that you can
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TPython
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Re: Very new to this - need help identifying if this is contam? [Re: Camera93] 1
#28688592 - 03/06/24 11:37 AM (3 months, 19 days ago) |
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Yeah don’t wipe the condiment cups down, they should be sterile out of the sleeve as they are made for food.
Count out and separate the amount of cups you’re going to be using for your agar session and remove them from the sleeve while still keeping them stacked together. Place them face down on your rack inside your SAB, do the same with the lids but keep them orientated topside up.
Then once you’re ready to pour you take a cup off the top of the stack turn it over upright just long enough to pour your agar and then take a lid off the top off the other stack and snap it closed and repeat.
Since the bottom most cup will have an air gap underneath it where contams can potentially get to it I never use it for agar but rather fill it with hot water and place it on top of the stack to help with condensation.
Hopefully that made sense. I’ll take some example pics if you’d like
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sasquanch
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Re: Very new to this - need help identifying if this is contam? [Re: joze]
#28688599 - 03/06/24 11:40 AM (3 months, 19 days ago) |
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Yep, I wiped down my SAB, wiped down the cups and lids and set them to the side, took out my agar in a media bottle that I PC at 15psi for 60 minutes. Waited till I could handle the jar (didn't have a laser thermometer yet) and poured into the cups and put the lid on asap. Stacked them up off to the side of the box and let them cool before resterilizing the box and introducing liquid culture.
I've got one of those popup plastic boxes that unzips on each side with the arm holes/sleeves. I don't like to synch the sleeves because it makes it hard to pull the syringe out to flame sterilize. Use a cooling rack on the inside to elevate the cups too.
I'm planning on making a purchase for some reusable / sterilizable agar plates but still looking for supplier that won't break the bank. Also looking for the best option for lids for the jars. Right now I'm just flipping the mason jar lid upside down for the gas exchange and adding foil. The first time I did the jars I didn't have a PC, so I just did the boil method for 90 min...those are having a hard time too. 3 of them look okay, but they are moving sloooowwwww. One looks kinda pink to me.
-------------------- sovereign.church
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sasquanch
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Re: Very new to this - need help identifying if this is contam? [Re: Smellyhobbit]
#28688602 - 03/06/24 11:42 AM (3 months, 19 days ago) |
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Sprayed the alcohol on a the cup and wiped down with paper towel and left to air dry off to the side
-------------------- sovereign.church
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Camera93
We got dicks like Jesus



Registered: 08/15/18
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Re: Very new to this - need help identifying if this is contam? [Re: sasquanch]
#28688603 - 03/06/24 11:42 AM (3 months, 19 days ago) |
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-------------------- All I need are some tasty waves, a cool buzz, and I’m fine. Whatever you decide won’t really impact our survival Close your eyes, and do the best that you can
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Very new to this - need help identifying if this is contam? [Re: sasquanch]
#28688611 - 03/06/24 11:49 AM (3 months, 19 days ago) |
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Quote:
sasquanch said: Yep, I wiped down my SAB, wiped down the cups and lids and set them to the side, took out my agar in a media bottle that I PC at 15psi for 60 minutes.
I only PC my agar for 30 minutes in an Instant Pot, you can save yourself a bit of time here.
Quote:
sasquanch said: Waited till I could handle the jar (didn't have a laser thermometer yet)
I use a typical glass thermometer. Once my PC has released pressure and is safe to open, I set the glass thermometer in the water and walk away. I periodically check the temperature, and swirl my bottle of agar to make sure the heat is distributing evenly. Once the temperature reaches 50°C, it's cool enough to handle, but it's still warm enough that it won't get chunky.
Quote:
sasquanch said: Right now I'm just flipping the mason jar lid upside down for the gas exchange and adding foil.
That's what I do. I love unmodified mason jar lids. I cover mine with foil when they're in the pressure cooker, but after that I remove the foil.
As we've already stated, you're making a mistake by wiping out your cups with alcohol. You should be either buying pre-sterilized cups, or switching to a no-pour TEK.
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sasquanch
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Re: Very new to this - need help identifying if this is contam? [Re: joze]
#28688665 - 03/06/24 12:37 PM (3 months, 19 days ago) |
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Awesome, I'll take that advice and fingers crossed I can salvage something from these current cups. I have a few that are growing something...just don't know yet what it is lmao
Appreciate the help!
-------------------- sovereign.church
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