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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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sigh, 2nd attempt at agar, do i just have a plate full of contams?
    #25845740 -

Hi guys,
Like the title says, this is my 2nd attempt at agar. Inoculated from MS Syringe 6 days ago. Growth started appearing by day 4. It kinda looks like the mould that grew in my previous agar attempt but whats weird is that my control plates are all negative for anything.



So whats the damage?

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845759 -

Transfer clean mycelium away from contaminants to a new plate. That is the whole point of agar.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845760 -

Looks like myc to me
Try transferring the leading edge away onto a new plate. a few time.


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when god hands you lemons you,FIND A NEW GOD! 

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: Skubydue]
    #25845786 -

Skubydue said:
Transfer clean mycelium away from contaminants to a new plate. That is the whole point of agar.


Yeh but I am not sure if I have any mycelium at all. My previous try I only had contams and they kinda looked like this but they had sporulated by this time and turned greenish.

If I can confirm that there are any good cubensis mycelia, I will isolate.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845795 -

They look like mycelium, but like u daid, penecilium also looks like that b4 it turns green. It should have started by now tho, I think it's safe to say it's mycelium. Penecilium grows in circles like that but usually the center would be green already.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: nube424]
    #25845800 -

I can’t see shit on dishes like those...


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Need a lift?

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: Hoob33]
    #25845816 -

i think you've got some cube myc growing. I'm guessing you just shot the load right onto the agar. I used to do this but have found streaking to be very beneficial. I recently started 7 new varieties from vendor syringes and they were all loaded with bacteria. But, thanks to the streaking tek, I was able to get clean (i think, time will tell) cultures with just one transfer.




And here they are a couple days later



If i hadn't streaked the agar, all that bacteria and myc would be growing together in a big pile. That'd making cleaning it up and identifying what is what a lot harder. In my opinion, when myc is piled up on itself or growing on top of bacteria, it looks a lot like what you've got there.

and yes, the fact that it hasn't turned green yet is a really good sign


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: BeefExtreme]
    #25845819 -

BeefSupremeJr said:
I can’t see shit on dishes like those...


Haha yeh sorry I'm not confident enough in working with open pour petris even in my SAB I guess my current confusion is a point in case.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: verum subsequentis]
    #25845846 -

:whathesaid:

And that definitely looks like mycelium, not bacteria or mold. I honestly have a really hard time growing any other mycelium than the mycelium I'm growing. Usually its bacteria and one time it was green mold and I think that was because of the mycelium syringe I used. It's all in the TEK. Streak it up homie!


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Back from my Hiatus :smile:

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: nube424]
    #25845851 -

nube424 said:
They look like mycelium, but like u daid, penecilium also looks like that b4 it turns green. It should have started by now tho, I think it's safe to say it's mycelium. Penecilium grows in circles like that but usually the center would be green already.


What's the usual time frame for sporulation with penicillium?

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845856 -

It's usually pretty fast. My oyster mushroom mycelium grew a lot faster though. Still used some samples from that tray and didn't get contams, but I also have a flow hood and faced the contam down wind.


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Back from my Hiatus :smile:

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: Jooceman]
    #25845863 -

And you don't worry about that spreading tons of mold spores around your lab?


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: verum subsequentis]
    #25845869 -

verum subsequentis said:
And you don't worry about that spreading tons of mold spores around your lab?


What?

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845877 -

it's a pretty straight forward question. He said he opens mold infested plates in front of his flow and I'm asking if he worries about the amount of spores that that is blowing all over his lab and house.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845879 -

I don't think penecilium moves fast like trich anyway... even if u had it on a plate and transferred to grain, I don't think it would fuck ur grow like bacteria or trich will. It's a pretty slow mold, I've noticed it almost stops growing after it sporulates. But what do I know.

After all, we've all eaten bread with penecilium in it anyways.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: verum subsequentis]
    #25845888 -

verum subsequentis said:
i think you've got some cube myc growing. I'm guessing you just shot the load right onto the agar. I used to do this but have found streaking to be very beneficial. I recently started 7 new varieties from vendor syringes and they were all loaded with bacteria. But, thanks to the streaking tek, I was able to get clean (i think, time will tell) cultures with just one transfer.




And here they are a couple days later



If i hadn't streaked the agar, all that bacteria and myc would be growing together in a big pile. That'd making cleaning it up and identifying what is what a lot harder. In my opinion, when myc is piled up on itself or growing on top of bacteria, it looks a lot like what you've got there.

and yes, the fact that it hasn't turned green yet is a really good sign



Thanks man, really nice looking plates. What media is that? Or is it just food coloring?

I'm familiar with streaking coming from a microbio background but always in a flow hood. Once i get at least one successful cultivation,  I'll be more willing to try some open air work albeit in the SAB.

Do you normally drop a small amount of spore solution and then streak or do you dip the loop into spore solution?

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845902 -

Small drop then streak. And a sab is almost just as good as a flow hood. Most people on here use sabs.

Another way is to slightly insert the needle into the agar and inject 1 or 2 drops INTO the agar, not on top. It'll hopefully keep bacteria below the surface and mycelium will surface and grow outward from the injection site. Just a little technique if used a few times. I prefer working with prints and swabs tho.

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: darkzen15]
    #25845918 -

Agar will favor contaminants, especially from a spore syringe.  You'll have much better luck with agar by using the zig zag motion mentioned above, but from a print rather than a syringe.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: RogerRabbit]
    #25845951 -

Hey roger, nice to see you man.

Darkzen, that's 2% Malt extract with a little food coloring (i think it's pretty). I flame my needle and my loop, shoot a tiny bit of solution out the needle (just a drop or two) to cool it, put one drop on the loop and then streak the plate.


--------------------
Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: sigh, 2nd attempt at agar, do i just have a plate full of contams? [Re: RogerRabbit]
    #25846006 -

RogerRabbit said:
Agar will favor contaminants, especially from a spore syringe.  You'll have much better luck with agar by using the zig zag motion mentioned above, but from a print rather than a syringe.
RR


Sorry I don't quite get what this means, you go directly from print to agar?

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