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Offlinetangra07
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Yellow on agar: contam?
    #26659971 - 05/09/20 12:47 PM (3 years, 9 months ago)

I have 2 agar jars, one of which has some yellow stuff on it, the other has only white spots. I am wondering, if the first one has a contam or if it is just agar, which got discolored somehow?

First one, with yellow spots:


Second one for comparison, only white:


Is the first one clearly contaminated and if yes with what? I used that same syringe on my last grow. I didn't notice any obvious contamination while colonizing, but it took super long for full colonization and after that it took super long to fruit and the harvest was really underwhelming, although the potency was superb.


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InvisibleRoger Clemency
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Re: Yellow on agar: contam? [Re: tangra07]
    #26660031 - 05/09/20 01:17 PM (3 years, 9 months ago)

It’s bacteria. Spore syringes often have some bacteria in them. Sometimes it’s all throughout the water it seems like and wherever it gets on the agar bacteria grows.

That’s probably why that grow had the issues you had. Those bacterial syringes seem to work pretty well with brf cakes for whatever reason but going spore syringe to grain often ends badly.


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Heaven starts right where hell ends


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Offlinetangra07
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Re: Yellow on agar: contam? [Re: Roger Clemency]
    #26660081 - 05/09/20 01:46 PM (3 years, 9 months ago)

Wow that explains a lot. It was grain. I made the error of innoculating 2 bags with the 2 strains and doing the agar after for fun. Now I see, that it would have actually helped me. Should I toss the bag I inoculated with the bad syringe? Its bird seeds, rice and line seeds. Or maybe just PC it again and inoculate again with a clean part from the agar?


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OfflineMachiavelliavore
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Re: Yellow on agar: contam? [Re: Roger Clemency]
    #26660082 - 05/09/20 01:48 PM (3 years, 9 months ago)

Maybe try sterilizing a q-tip in some foil (see eatyualive sterile swabs now) putting a drop of that syringe, and streaking.  Make sure there is not standing water on your plates, put'm upside down or or tap and dump the water.  That will get the bacteria everywhere.

You could also try going to a bit of grain or PF and transferring anywhere you get mycellium to agar.  Bacteria really thrives in wet conditions, so you might get mycellium on top of your grain and bacteria at the bottom.  Useless for spawn, but you can transfer those cleaner grains on top to agar.

If you do get any growth on those bacterial plates, there was an old method to pour agar over the mycellium to to isolate bacteria, but I would try placing a wedge from a clean plate on top of the mycellium, then picking it off the dirty plate once you have mycellial growth, and puttin it onto a clean plate.

I would use your bags to looks for grains with growth on them to transfer to agar.


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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Offlinetangra07
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Re: Yellow on agar: contam? [Re: Machiavelliavore]
    #26660191 - 05/09/20 02:42 PM (3 years, 9 months ago)

Sounds like an interesting exercise. What if I alcohol spray the agar jar? After all alcohol kills bacteria, but isn't very effective against fungi? Or if I cut a clean looking mycelium piece of the agar, put it on new agar, wait to see if its contam'ed, repeat until I see completely clean mycelium?

Im not sure I understood the method with pouring agar over the current agar. The idea is, that the bacteria won't travel up the new top layer agar, but the mycelium will?


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OfflineMachiavelliavore
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Re: Yellow on agar: contam? [Re: tangra07]
    #26660220 - 05/09/20 03:01 PM (3 years, 9 months ago)

Quote:

tangra07 said:
Im not sure I understood the method with pouring agar over the current agar. The idea is, that the bacteria won't travel up the new top layer agar, but the mycelium will?



Yeah, you have to pour it at a certain temperature, right above the gel point, I'm assuming like 110 or 105F.

I'm not really sure what the alcohol would do, prolly kill pretty much everything, I doubt it would be productive.  It's doubtful any mycellium will even develop on those plates, as it you would be asking for spore germination in a pool of bacteria.


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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Offlineincogneato
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Registered: 03/15/19
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Re: Yellow on agar: contam? [Re: tangra07]
    #26660549 - 05/09/20 05:59 PM (3 years, 9 months ago)

You've got good info here already. I'll just chime in to say those both look bacterial to me. Bacteria come in many colors. The most common one that I see personally looks just like that milky white stuff in both of your jars, if I'm not mistaken. It usually shows up right away, days before you'll see any mycelium start up. If you're trying to get a culture, wait for the mycelium to grow, then transfer an edge that isn't touching the bacteria to a new plate. Repeat until you get a clean culture.

If you're seeing a lot of bacteria only around the outside edges of your plate, it could be your handling technique rather than the material you're adding. But I agree about spore syringes. Not always clean.


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