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Mycopsilo
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What is this Contam on my Agar?
#26644063 - 05/02/20 03:52 PM (3 years, 8 months ago) |
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First Agar attempt. Plates are 8 days old.All work was done in an SAB. When I poured the plates I poured at 125F. I screwed up I guess because all the plates got condensation. I let the plates cool in the SAB. I inoculated 5 out of the 20 plates with a B+ MS syringe at 2 drops per plate. I sealed all the plates with plastic wrap. The extra plates i poured went to the fridge for storage. My inoculated plates have been in my lab between 70 and 80F for 8 days. 1 plate looks like it has 3 small circles of nice myc . Other 4 have some myc and some grayish raised growth circles all around the plates. they still have condensation as well.
What could these contams be? They look similar to the texture of the agar but raised circles and grayish color.Could they be from the condensation water? They are everywhere. What could have caused it? I flame sterilized my syringe as well. Plates weren't open for more than 3 seconds. My Agar is made of Nutritional Yeast,AGAR and barley malt. What should I do with the good plate? How do I upload a pic from my phone to post?
Edited by Mycopsilo (10/25/20 07:04 PM)
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Roger Clemency
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26646166 - 05/03/20 02:30 PM (3 years, 8 months ago) |
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Click on PICS under your name near the top left. Then click upload and it should take you from there. Once you upload to your gallery you can copy the URL and put it in your post.
What you’re explaining sounds like bacteria, probably from the syringe itself.
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mushpunx
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Spore syringes are never 100% clean. Neither are prints (and syringes are made from prints).
This is the reason *why* we use agar, to transfer clean myc away from contams. It was probably your syringe, unless your technique was sloppy and you introduced it that way. Post a photo, sounds like bacteria like said above. We don't really need to identify contams, it's always gonna be bacteria or mold.
With spore solution , after you place a drpp on the plate you should use a flame sterilized loop to swipe the spores across the plate in a zig zag pattern (loop up images of swiped plates). This makes it easier to identify clean germination.
Even if you think you have no contams, you should never use a germination plate anyways. Always make at least one transfer before putting to grains.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx] 1
#26646604 - 05/03/20 05:42 PM (3 years, 8 months ago) |
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: Mycopsilo] 1
#26646608 - 05/03/20 05:45 PM (3 years, 8 months ago) |
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26646613 - 05/03/20 05:46 PM (3 years, 8 months ago) |
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First one looks crazy. The second one looks good but they are connected am I right?
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26650333 - 05/05/20 10:37 AM (3 years, 8 months ago) |
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Does the second plate look good enough to transfer?
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Roger Clemency
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26650887 - 05/05/20 02:03 PM (3 years, 8 months ago) |
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Yeah you can transfer from either. The first one would be tougher of course but even with that second one it looks like there might be other stuff going on so either way you will want to do another 1 or 2 transfers after getting off those plates.
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panne cyanne
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bacteria. that plate is cooked.
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mushpunx
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Re: What is this Contam on my Agar? [Re: panne cyanne]
#26651397 - 05/05/20 06:23 PM (3 years, 8 months ago) |
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Holy shit that first plate is toast. You can see the myc, but it's grown over top bacteria- I would not bother to transfer from that.
The second one, take a transfer from that and see how it grows.
You really should be swiping the spores in a zig zag pattern across the plate with an inoculation loop when you inoculate.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26652124 - 05/06/20 03:47 AM (3 years, 8 months ago) |
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Do you think the condensation I had caused that bacteria? The whole lid was covered but I saw a post that said flip the plates upside down to get rid of the condensation. It worked but there was still some residual water.. Is there any other way to get the condensation out if I screw up another batch of agar?
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26652149 - 05/06/20 04:03 AM (3 years, 8 months ago) |
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Quote:
Mycopsilo said: Do you think the condensation I had caused that bacteria? The whole lid was covered but I saw a post that said flip the plates upside down to get rid of the condensation. It worked but there was still some residual water.. Is there any other way to get the condensation out if I screw up another batch of agar?
No, the condensation doesn't cause bacteria. I wait until the agar is cool to the point it might start solidifying if I let it go any longer before I pour, and I never get condensation. If you do, you can try setting a mug of hot water on top of the plate and let it cool.
Unless you have sloppy technique, and exposed it to contams that way, it was most likely in your spore syringe. No print or the spore syringes we make from them are ever 100% clean. That is why we use agar in the first place.
But by just placing a drop of spore solution on the plate the spores are going to germinate in that one area along with whatever bacteria or contams too. See how in the 1st plate the myc has grown overtop the bacteria? Sometimes the myc can hide bacteria or mold like that. That is why it's best to place the drop of spore solution (or swipe a print) on the plate and then swipe it in a zig zag pattern across the plate. (Look up photos of swiped plates). This will make it easier to get clean myc away from contams, and you should know roughly where the germination is occuring.
Mold tends to appear as little white colonies on the plate that can resemble mushroom myc , so you should avoid anything outside the zig zag pattern.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26653505 - 05/06/20 06:28 PM (3 years, 8 months ago) |
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I took two transfers from the good plate. How long do you think until the new plates are colonized? I’m already seeing myc expanding to the new agar.
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26653969 - 05/06/20 10:05 PM (3 years, 8 months ago) |
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Quote:
Mycopsilo said: I took two transfers from the good plate. How long do you think until the new plates are colonized? I’m already seeing myc expanding to the new agar.
A few days usually. I never let them grow out more than 3/4 or so before putting them into cold storage. Might not hurt to take yours out to T3 though.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26654535 - 05/07/20 07:12 AM (3 years, 8 months ago) |
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What is the optimal temperature for cubes to colonize? What is the optimal temp for fruiting?Do most cubes generally require the same temps? I see mixed posts and I just want a straight answer. I’ve got Albino A+, B+, PE, GT and Treasure Coast.
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26655076 - 05/07/20 12:36 PM (3 years, 8 months ago) |
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Quote:
Mycopsilo said: What is the optimal temperature for cubes to colonize? What is the optimal temp for fruiting?Do most cubes generally require the same temps? I see mixed posts and I just want a straight answer. I’ve got Albino A+, B+, PE, GT and Treasure Coast.
I don't usually worry about temps too much. Cubes are a hardy creature. 60 and up is fine for colonization , but higher temperatures during spawn colonization might encourage bacteria (Like high 80's, 90's). As far as post spawning goes, I've fruited Cubes in a hot stuff attic before, it would get as low as 50's at night that would swing well above 100F during the day - took a lot to keep them fr drying out but they did just fine. Higher temps might cause some hollow fruits, low temps more solid.
So yeah if I had to pick an optimal temp it would be 70F. Most of us don't worry about it.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26655499 - 05/07/20 04:18 PM (3 years, 8 months ago) |
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That’s what I was thinking. My room is fluctuating between 63 at night to a little more than 70 during the day.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26655652 - 05/07/20 05:38 PM (3 years, 8 months ago) |
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So after my plates are done I plan on spawning to rye in quart jars. Do you think 2 rice size pieces of agar is enough to colonize each jar? Im guessing it is but the more pieces the faster the colonization I would think. I will be inoculating 7 jars total then spawning to coir only bulk substrate. I would like a 1:2 spawn to substrate ratio. I’m using a 66QT Monotub and I’m wondering how much coir I’m going to need for a four inch substrate depth. Is that enough spawn to achieve that ratio? I tried using the substrate calculator but I’m not sure it factors the coir when it is hydrated to field capacity. I don’t know how much the coir expands when hydrated. I don’t want to waste coir even though it’s relatively cheap. I realize I’m going to need to experience some trial and error to learn but if you could guide me a little it could save me from some of the error. Thanks again for your help, it is much appreciated.
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26655762 - 05/07/20 06:32 PM (3 years, 8 months ago) |
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Rice sized? No. You can use dime-nickel size pieces. Personally I use a quarter plate per jar, it colonizes faster.
Typically I use an entire brick of coir per tub. I will use 4 to 6 quarts per tub. You could use 7 quarts to one tub, it will just mean a higher yield.
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LogicaL Chaos
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26655766 - 05/07/20 06:34 PM (3 years, 8 months ago) |
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Quote:
Mycopsilo said:

The slimy stuff is bacteria, the white is well mycelium (in case u didnt know )
Also, awesome picture!
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Mycopsilo
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Thanks, I have gathered that much information Lmao.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26656220 - 05/07/20 10:23 PM (3 years, 8 months ago) |
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Got it. Thanks. I will be updating my progress and posting pics as it goes.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: Mycopsilo] 1
#26667001 - 05/12/20 04:23 PM (3 years, 8 months ago) |
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Here is the B+ transfers. I wasn’t thinking when I did it and took the myc from the center... They are looking good though from what I can tell. The first plate I dropped received two pieces of agar by accident. A little sliver fell off of the main transfer. The second plate got a single piece. The second plate has a small dot of what looks like the start of some new myc?

Edited by Mycopsilo (05/12/20 04:31 PM)
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26667940 - 05/13/20 04:37 AM (3 years, 8 months ago) |
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The second plate, that dot could either juSt be a piece of myc that fell off your scalpel or if your wedge rolled, and started growing.
But it also could be a mold colony - mold colonies start out as white spots like that, in places not touching your wedge or myc germination. That's why we swipe spores onto plates in zig zag patterns, so we have an idea where germination should be and don't take from outside of the pattern.
Plates like that I view as suspect, I take a transfer from the side furthest from the colony, before the myc touches it. That's what I would do.
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26668086 - 05/13/20 06:52 AM (3 years, 8 months ago) |
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Ok that’s what I’ll do. On a similar note I made some new Agar plates and did some swipes from my syringes. 5 more B+ and 5 PE. I poured the agar at 120F and I still got condensation.. What the hell is up with that? Every post I read said it starts to solidify at 120F but that wasn’t the case. So I noticed the agar was still liquified at 100F so I guess I will pour at 100 from now on. I don’t think My IR thermometer is off.
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mushpunx
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Re: What is this Contam on my Agar? [Re: Mycopsilo]
#26668152 - 05/13/20 07:50 AM (3 years, 8 months ago) |
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You could go cooler if it's still causing problems. I eyeball mine a lot of the time, when it's just about going to solidify if I leave it any longer, I pour. Never get condensation.
I usually only do one or two germination plates of each variety because I'm just gonna looking to start one one culture of each. Saves plates
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Mycopsilo
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Re: What is this Contam on my Agar? [Re: mushpunx]
#26668198 - 05/13/20 08:19 AM (3 years, 8 months ago) |
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Ok. So you you just transfer off of those 2 plates to make more plates? How do you isolate? You just transfer off until you have a culture that is completely uniform?
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