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OfflineBIOMA
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Registered: 04/14/20
Posts: 60
Last seen: 2 years, 5 months
Self made PDA agar * 1
    #26670117 - 05/14/20 07:05 AM (3 years, 8 months ago)

Hello guys!

I'm about to start working with agar for reducing contaminants and create proper LC to inoculate my jars that I have been doing using the PF TEK.

So I have a couple questions:

1) From what I have read, PDA agar is the best for this. Unfortunately is not possible to buy it in the place where I am right now. So I bought Agar Agar, Dextrose and potatoes and I'm going to try and do PDA myself following some recipes on the internet. Anybody have experience with this? does it work? any tips?

2) My main purpose is to do as many transfers as I have to, in order to remove contaminants for a posterior inoculation. How can I take the mycelium from the Petri dish and put id inside of a syringe or a jar to create the liquid culture (LC) or what's the technique for inoculating PF TEK jars from mycelium in agar?

Please help me, this is holding me back from advancing since all my experiments with spore water have failed because of contaminants and I don't see any other way to avoid this than the magic of the agar.

Thank you ?


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NATURE IS EVERYTHING AND EVERYTHING IS NATURAL



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OfflinePinkStormtrooper
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Registered: 04/11/20
Posts: 218
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Re: Self made PDA agar [Re: BIOMA]
    #26670139 - 05/14/20 07:32 AM (3 years, 8 months ago)

you can boil potato and go that way if you have some time to kill, but just go scrum up some potato flakes, or dog food, or cat food, or fish food, find some protiens man


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"say, you got a little astroglide on your moustache"


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InvisibleStrainsfordaze


Registered: 05/10/18
Posts: 669
Re: Self made PDA agar [Re: BIOMA]
    #26670141 - 05/14/20 07:32 AM (3 years, 8 months ago)

1. Potatoes, agar and dextrose will work out fine.

2. With a scalpel in a SAB or in front of a flow hood.
You could inoculate with the LC syringe, but at that point you could be progressing beyond PF tek to grain prep.

Agar>LC>grains>bulk.

There are many ways to achieve success. 

You could go agar>grains>bulk and skip the LC step.


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OfflineCamera93
We got dicks like Jesus
I'm a teapot


Registered: 08/15/18
Posts: 3,220
Last seen: 10 days, 11 hours
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Re: Self made PDA agar [Re: BIOMA]
    #26670145 - 05/14/20 07:33 AM (3 years, 8 months ago)

Here is a good write up for using no pour agar. It has the recipe for PDA. I have used this with success

pastys ez agar tek

This is another write up showing you what to look for as far as making transfers and getting a clean culture

C10's cleanin up on agar tek

for Liquid culture

mushboys write up on LC


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All I need are some tasty waves, a cool buzz, and I’m fine.

Whatever you decide won’t really impact our survival
Close your eyes, and do the best that you can



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OfflineBIOMA
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Registered: 04/14/20
Posts: 60
Last seen: 2 years, 5 months
Re: Self made PDA agar [Re: Strainsfordaze]
    #26676468 - 05/17/20 08:32 AM (3 years, 8 months ago)

OK cool, thanks :smile:


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NATURE IS EVERYTHING AND EVERYTHING IS NATURAL



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OfflineBIOMA
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Registered: 04/14/20
Posts: 60
Last seen: 2 years, 5 months
Re: Self made PDA agar [Re: Strainsfordaze]
    #26676477 - 05/17/20 08:36 AM (3 years, 8 months ago)

Thank you, you are right, but Im in a learning process, thats why I want to master the PF tek before moving to grain. But I find agar as the only solution no to contaminate the substrate... all my spore prints seem to be not clean enough.

SoI know Its possible to inoculate with the LC, but my question is what's the safest way to transfer mycelium from agar to water to create LC syringes.


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NATURE IS EVERYTHING AND EVERYTHING IS NATURAL



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OfflineBIOMA
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Registered: 04/14/20
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Last seen: 2 years, 5 months
Re: Self made PDA agar [Re: Camera93]
    #26676478 - 05/17/20 08:36 AM (3 years, 8 months ago)

This is awesome! Thank you :smile:


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NATURE IS EVERYTHING AND EVERYTHING IS NATURAL



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OfflineSockadin
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Registered: 01/03/10
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Re: Self made PDA agar [Re: BIOMA]
    #26676769 - 05/17/20 12:01 PM (3 years, 8 months ago)

So after you have gotten a good amount of clean agar culture. Take a small wedge and drop it into a grain water LC. Let that grow out for a week and suck up the water into the syringe. You can make a self healing injection port in your lid by drilling a 1/4 hole and covering in RTV or Silicone. Then stick your needle in the hole to retrieve the culture when it has grown out.

Your gonna need a PC for all of this and might as well go to grain at that point.


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Offlinemushpunx
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Re: Self made PDA agar [Re: Sockadin]
    #26677066 - 05/17/20 02:52 PM (3 years, 8 months ago)

BRF is pretty forgiving when it comes to spore syringes, if you are getting lots of contamination you might want to work on your syringe making technique. Shouldn't be getting many contams for Pf tek

Good move going to agar! Honestly though, you should start with grains. Learn to inoculate grains with agar wedge first, G2G, etc.


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