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Offlinekspohr
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1st attempt at agar!
    #26877724 - 08/12/20 11:53 PM (3 years, 6 months ago)

potato starch / dex / agar / yeast

PC'd for 30mins 15psi

PE #6 spores

day 10.











how does it look to everyone?
im not sure if those pink dots are lipstick mold or not.
not sure if there is to much moisture in there.

the second last photo looks like it has a contaminate that needs to be removed.


Edited by kspohr (08/12/20 11:54 PM)


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InvisibleGoatrider
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Re: 1st attempt at agar! [Re: kspohr]
    #26879589 - 08/14/20 06:06 AM (3 years, 6 months ago)

I`d send these to hell.
That`s bacteria and mold.
Looks dangerous.
What was the colour like before inoculation?
You have to improve sterile techniques.

Have a look at the link:
https://www.shroomery.org/forums/showflat.php/Number/24330957


                        :cookiemonster:


Edited by Goatrider (08/14/20 06:17 AM)


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Offlinekspohr
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Re: 1st attempt at agar! [Re: kspohr]
    #26880535 - 08/14/20 06:16 PM (3 years, 5 months ago)

The color before inoculation for the agar is that deep purple color.

I am starting to wonder if the spore syringe i have is no good. This is the second batch from the same Spore Syringe. First one batch came out with lots of pink dots as well, didnt make it as far as these current ones. I threw them out before anything white appeared.

I have read a bunch of bodhi's posts.
My agar was done similar to Pastywhyte's Easy Agar Tek different color and PDA + yeast. This was pressure cooked @15psi for 30min.

I have made a SAB with 3" holes with gloves attached and put this SAB in my small den (4x8) room.
Before I used my SAB, I would go in and spray the air down and wiped the desk, SAB, PC, lighter, spore syringe etc.
Everything was wiped down with lysol wipes or 75% iso.
Sprayed myself before i would enter the room after i cleaned it up as well and had gloves on along with a mask.
All of this was done before inoculating.


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InvisibleGoatrider
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Re: 1st attempt at agar! [Re: kspohr]
    #26880893 - 08/14/20 11:47 PM (3 years, 5 months ago)

Perhaps we have the answer now.
I would advice to remove the attached gloves,
that`s outdated, but can work perfect if you know exactly what you`re doing,
let`s say for G2G, as you don`t have to move out, once loaded your SAB.
Air pressure may change due to your movements in the gloves,
and forces airflow in.
A SAB never has gloves attached, that`s a glove box,
and it totally destroys the purpose of a SAB.

I remember also being a bit paranoid, as i started with a SAB.
Most important are your evenly movements, don`t overdo with too much
sanitizers. Close doors and windows, spray room down a bit, clean the working surface, perhaps a bit soapy water to the SAB and you`re good to go.


                              :cookiemonster:


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Offlinesmalltalk_canceled
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Re: 1st attempt at agar! [Re: Goatrider]
    #26880901 - 08/14/20 11:56 PM (3 years, 5 months ago)

Its probably the syringe.


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InvisibleGoatrider
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Re: 1st attempt at agar! [Re: smalltalk_canceled]
    #26880903 - 08/14/20 11:59 PM (3 years, 5 months ago)

Of course that`s also in the game.


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OfflineOne of Us
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Re: 1st attempt at agar! [Re: Goatrider]
    #26880906 - 08/15/20 12:04 AM (3 years, 5 months ago)

It looks like you can grab something from the 6 o clock area of plate 3

Using purple agar always looks interesting because most of the time, the mycelium will eat the blue dye and leave behind the red. This is what is happening in your case


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Invisiblealaskappalachian
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Re: 1st attempt at agar! [Re: One of Us]
    #26880922 - 08/15/20 12:34 AM (3 years, 5 months ago)

^^^ This, and those plates are loaded with bacteria.  Next time just use a DROP.  If you're using a syringe, consider wrapping some qtips in foil and throwing them in a jar to sterilize for like 20min @15psi to make some makeshift swabs to put a little solution on to swab the plate.  All that spore solution makes for a bacterial mess (all syringes have some contamination so more solution introduces more contaminants).  You can also make an inoculation loop from the metallic inside a twist tie for garbage bags to hold a drop of solution.  :thumbup:


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InvisibleGoatrider
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Re: 1st attempt at agar! [Re: One of Us]
    #26880923 - 08/15/20 12:35 AM (3 years, 5 months ago)

Quote:

One of Us said:
It looks like you can grab something from the 6 o clock area of plate 3





Yeah, could be worth a try, if you really need the plates.
I´d just start a new batch, ideally with a spore print.


                        :cookiemonster:


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OfflineOne of Us
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Re: 1st attempt at agar! [Re: Goatrider]
    #26880934 - 08/15/20 12:43 AM (3 years, 5 months ago)

Yeah, I just noticed those pics were posted 2 days ago, so it is likely not salvageable anymore anyway


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Invisiblec10h12n2o
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Re: 1st attempt at agar! [Re: One of Us]
    #26880937 - 08/15/20 12:54 AM (3 years, 5 months ago)

Quote:

One of Us said:
It looks like you can grab something from the 6 o clock area of plate 3

Using purple agar always looks interesting because most of the time, the mycelium will eat the blue dye and leave behind the red. This is what is happening in your case




Looks like it has bacteria even on the part the myc hasnt touched yet

Id toss all that

@op did you do any control plates? You should. Thatll help you start your debugging. If those are good,  try streaking the plates like the gif in my sig (explained in agar guide)

Even if the syringe is dirty, you should be able to get clean myc pretty easy by using serial streaking. Just focus on your technique


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Edited by c10h12n2o (08/15/20 12:55 AM)


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InvisibleLotKid
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Re: 1st attempt at agar! [Re: c10h12n2o]
    #26881129 - 08/15/20 06:43 AM (3 years, 5 months ago)

Also... opening them to get a pic of them... not a good idea.


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Offlinekspohr
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Re: 1st attempt at agar! [Re: kspohr]
    #26882453 - 08/15/20 10:19 PM (3 years, 5 months ago)

thanks for the help everyone.

I already threw everything out and I've already started a new set of agar dishes.
I'm going to try the streaking on the dish this time.
I have another spore syringe from this supplier, i hope its my process. I know where i need to make corrections in my process. Hopefully its enough to make things work.

I had to much moisture in the agar to begin with, in combination with to much from the spore syringe.

also what does he mean by control plates?


Edited by kspohr (08/15/20 10:57 PM)


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Invisiblec10h12n2o
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Re: 1st attempt at agar! [Re: kspohr]
    #26882539 - 08/16/20 12:02 AM (3 years, 5 months ago)

Take a sterile plate and dont inoculate it or open it.  Take another plate and open it in your sab , then close it without inoculating

Then procede with inoculation of a 3rd plate

Those 2 control plates will give you a good idea which point things are going sideways


--------------------

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche


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