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Offlinemaxime59
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First time agar grower
    #26491932 - 02/18/20 03:06 PM (3 years, 11 months ago)

Hello !

First time posting on Shroomery after a lot of stalking :cool:

I'm a new grower and I'd like to share with you my experience including a few pictures of my first plates.

My mushroom growing background is null. I used to grow mushrooms from growing kits and recently decided to go all-in from MS to agar to grains.

Being not much confident about the procedure, I've practiced, quite a lot, parts of the workflow:

  • Using my pressure cooker
  • Cleaning & working inside my SAB
  • Wrapping petri dishes
  • Sterelizing glass petri dishes (even though I still don't feel comfortable with them compare to the plastic ones)
  • ...

I've decided to inoculate many plates (maybe too much) expecting a lot of them to go wrong in the process.

Here's list of things that might need improvements in my workflow:

  • I feel like my agar solution doesn't look clear enough. Too much sediments end up in my plates. This brings confusion when attempting to recognize healthy mycelium.
  • I have a hard time maintaining the 15-17psi range during sterilization. 20 mins sterilization time can be managed (i.e. for agar) but will be unbearable for items that need more time (e.g. grains jars)
  • Looking at videos on Youtube, it looks like I'm too much paranoid: using too much gloves, clean towels, isopropyl alcohol, wasting a lot of time cleaning my working area ... (this will cost me $$)
  • I've yet to see something that looks like contams in order to improve my identification & isolation skills.
  • I'm not much comfortable with my glass petri dishes sterilizing technique: clean them with soap and warm water, wrap them in foil and put them inside my oven for 30 mins at 350F.

Please feel free to guide me on these few points :smile:

My plates are inside a box in a medium sized living room with natural ambient light. I'm able to maintain 72F (inside the box) by heating the room (it's winter time)

Here's a few pictures of my 1st transfer (receiving plates). Please bear in mind I'm a beginner, I've yet to learn how to take good pictures that favor contams identification.



Let me know if you see something wrong in them :smile: I've decided to proceed with a 2nd transfer but feel like I could have used them directly to inoculate my grains.

Looking forward to hearing from you'all.

Peace!


Edited by maxime59 (02/18/20 03:09 PM)


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Offlinepsycho_nauticus
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Re: First time agar grower [Re: maxime59]
    #26491955 - 02/18/20 03:22 PM (3 years, 11 months ago)

Holy crap, first time?!?!? Dude I wish I had plates like that for my first time. I'm still waiting to see how my agar inoculation turns out, and I'm new at it also.


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OfflineMr.Wizard
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Re: First time agar grower [Re: psycho_nauticus]
    #26492004 - 02/18/20 03:52 PM (3 years, 11 months ago)

I just started too, my sediment issues were solved when I made grain water agar:



One thing you can do is look from the bottom with light from the top, anything that is sediment should not cast any kind of shadow. On the reverse you can check if it's sediment by flipping the dish, and seeing if it casts a shadow from the bottom.

Here's my firsts (one transfer from germ plate):



And current:



--------------------
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InvisibleLadysKnight
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Re: First time agar grower [Re: Mr.Wizard]
    #26492051 - 02/18/20 04:35 PM (3 years, 11 months ago)

@maxime59 - 2nd plate, 3rd pic. See the gray wispy right half of the plate, that's mold. If you look at most of the others, you'll see it there too. The first one could be clean, I'd take more transfers.

On sterilizing glass petris, I find it easier to pour the agar, wrap a stack, then sterilize them in the PC. Ready to use when cool.


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Offlinejbgtaa
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Re: First time agar grower [Re: LadysKnight]
    #26492067 - 02/18/20 04:43 PM (3 years, 11 months ago)

Your agar looks fine OP. Some of those plates are either contaminated or just not isolated enough but there are good transferable sections on all of them. Also it’s clear enough. Idk what medium your using but malt extract will always be a little opaque.


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Offlinemaxime59
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Re: First time agar grower [Re: LadysKnight]
    #26492164 - 02/18/20 05:29 PM (3 years, 11 months ago)

Thank you all for the feedback ! :smile:

@jbgtaa: I use BRF agar (1.5%). Let me clarify, this medium troubles me mainly because I have big sediment spots on some of my plates. Being new to this practice, it's quite challenging differentiate them from actual contams.

@Mr.Wizard: Great work ! Thanks for the tips. I'll take a look at grain water agar.

Quote:

LadysKnight said:
@maxime59 - 2nd plate, 3rd pic. See the gray wispy right half of the plate, that's mold. If you look at most of the others, you'll see it there too. The first one could be clean, I'd take more transfers.

On sterilizing glass petris, I find it easier to pour the agar, wrap a stack, then sterilize them in the PC. Ready to use when cool.




Quote:

LadysKnight said:
Your agar looks fine OP. Some of those plates are either contaminated or just not isolated enough but there are good transferable sections on all of them. ...





Wow. I'm grateful to you guys ! I kept having this idea of mold looking way more darker than that. I'll try to post an update of my 2nd transfer in a few days.


Edited by maxime59 (02/18/20 05:30 PM)


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OfflineBackbone
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Re: First time agar grower [Re: maxime59]
    #26492231 - 02/18/20 06:05 PM (3 years, 11 months ago)

Those look beautiful but if youre cloning what is the point


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Offlinemaxime59
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Re: First time agar grower [Re: Backbone]
    #26501307 - 02/24/20 11:14 AM (3 years, 10 months ago)

Hello again,

Here's two plates of my 2nd transfer after 6 days, extracted from the red plates in my first post.



I went for a 3rd transfer as I still see this gray (not so white) color in the middle. What do you think ?

Thanks in advance for your help.


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OfflineA.k.aM
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Re: First time agar grower [Re: maxime59]
    #26501314 - 02/24/20 11:17 AM (3 years, 10 months ago)

That looks good to me man, I think the grey in the middle is probably from the myc eating up the coloring.


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LAGM2020


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Invisiblebucket
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Re: First time agar grower [Re: maxime59]
    #26513055 - 03/02/20 03:21 PM (3 years, 10 months ago)

Quote:

maxime59 said:
Hello again,

Here's two plates of my 2nd transfer after 6 days, extracted from the red plates in my first post.



I went for a 3rd transfer as I still see this gray (not so white) color in the middle. What do you think ?

Thanks in advance for your help.



Quote:

maxime59 said:
Hello again,

Here's two plates of my 2nd transfer after 6 days, extracted from the red plates in my first post.



I went for a 3rd transfer as I still see this gray (not so white) color in the middle. What do you think?

Thanks in advance for your help.




This is what I was wondering, too... how many transfers to expect when starting with MS Syringe to agar.
So, from there, the point of more plates would be to have more for each grain jar?


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OfflineA.k.aM
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Re: First time agar grower [Re: bucket]
    #26513061 - 03/02/20 03:25 PM (3 years, 10 months ago)

When I started I was making a plate per jar, then realized g2g is way better.

Doesn’t hurt to have plenty of backups of a culture though.


--------------------
LAGM2020


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Offlinepsycho_nauticus
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Re: First time agar grower [Re: A.k.a]
    #26513117 - 03/02/20 03:52 PM (3 years, 10 months ago)

So how much do you use for a jar? A fourth piece? So a dish can inoculate 4 jars?


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Offlinemaxime59
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Re: First time agar grower [Re: psycho_nauticus]
    #26513132 - 03/02/20 04:05 PM (3 years, 10 months ago)

Quote:

psycho_nauticus said:
So how much do you use for a jar? A fourth piece? So a dish can inoculate 4 jars?




I've used theses two plates to inoculate 8 jars using 1/4 piece per jar. I also decided to inoculate 2 more extra jars using another backup plate that I had (expecting at least one of them to go wrong) in order to attempt a g2g transfer inside my SAB later on.

It might sound unnecessary to do that much but being new to this, I make an effort to not put all my eggs in one basket. That is, I'll perform both (a2g & g2g) and see what happens.


Edited by maxime59 (03/02/20 04:12 PM)


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OfflineA.k.aM
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Re: First time agar grower [Re: maxime59]
    #26513158 - 03/02/20 04:21 PM (3 years, 10 months ago)

I’m impatient the least I’ll use is half a full size plate. A couple times I had stacks of clean and  plates for a clone and put two whole ones in a quart. Really any amount will work. That’s another benefit of soft plates,  they’ll break up when you shake them and cover more grains.


--------------------
LAGM2020


Edited by A.k.a (03/02/20 04:22 PM)


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OfflineBackbone
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Re: First time agar grower [Re: A.k.a]
    #26513228 - 03/02/20 04:56 PM (3 years, 10 months ago)

By the time your agar grows fully on your plate you could make up for the difference in a grain jar


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OfflineA.k.aM
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Re: First time agar grower [Re: Backbone]
    #26513426 - 03/02/20 06:51 PM (3 years, 10 months ago)

Oh for sure I just went overboard on a couple clones and had like 6 grown in plates sitting around. I’m all about the g2g now


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Offlineantmanmax
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Re: First time agar grower [Re: A.k.a]
    #26513459 - 03/02/20 07:09 PM (3 years, 10 months ago)

Beautiful! You've gotten sterile technique down, and you've gotten transfers down. Amazing first attempt (and it seems you got lucky with your spores as well).

Just remember to chill out. You're able to admit you're paranoid, which is good, but there are some aspects of this hobby where paranoia can fuck things up real quick.


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Offlinemaxime59
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Re: First time agar grower [Re: antmanmax]
    #26539192 - 03/16/20 06:43 PM (3 years, 10 months ago)

Thanks again to you all for the feedback :smile: Here's a little update :

I've done the A2G transfer like described previously. The colonization seems to go well.


I've already shake them. Twice, for some of them as it was a hard to break/distribute the grains by hand. However, in a few of my jars, we can see that the colonized grains weren't well distributed after shaking. Resulting in big "empty" areas (yet to be colonized?).


I don't feel comfortable to shake them more than twice as I've read that it was counterproductive. What do you think I should do ?

Another question : Two of my jars look fully colonized from the outside. Should I put them in the fridge in order to avoid pinning (waiting for the other jars to be done) ?

Thanks in advance for you help.


Edited by maxime59 (03/16/20 06:47 PM)


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InvisibleLadysKnight
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Re: First time agar grower [Re: maxime59]
    #26539213 - 03/16/20 06:58 PM (3 years, 10 months ago)

Quote:

maxime59 said:
Thanks again to you all for the feedback :smile: Here's a little update :

I've done the A2G transfer like described previously. The colonization seems to go well.


I've already shake them. Twice, for some of them as it was a hard to break/distribute the grains by hand. However, in a few of my jars, we can see that the colonized grains weren't well distributed after shaking. Resulting in big "empty" areas (yet to be colonized?).


I don't feel comfortable to shake them more than twice as I've read that it was counterproductive. What do you think I should do ?

Another question : Two of my jars look fully colonized from the outside. Should I put them in the fridge in order to avoid pinning (waiting for the other jars to be done) ?

Thanks in advance for you help.




I shake mine after inoc, because it's also the shake to distribute the moisture after PC (wbs).

I shake once more around 40%.


If it's not recovering well after that second shake, I know it's a loss, usually too bacterial.


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OfflineA.k.aM
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Re: First time agar grower [Re: LadysKnight]
    #26539221 - 03/16/20 07:03 PM (3 years, 10 months ago)

There’s sort of a technique to shaking so the grains mix well.  I tilt the jar an inch or two and then smack it five or six times real fast and it rotates all the grains so the bottom ones end up on top. If you do it a couple times rotating the jar a little bit in between they’ll mix up well.

As long as they’re healthy they’ll get to the bare parts super fast at the end.

It looks like you have very slight bacteria based on some of the thick rhizo but definitely not enough to be a problem with how much is already colonized and looks healthy.


--------------------
LAGM2020


Edited by A.k.a (03/16/20 07:05 PM)


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