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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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How do you (personally) prove your isolates?
    #12296397 -

With this being my first attempt at dabbling in the world of agar, and strain isolation's,( still waiting on all materials to arrive) just curious as to how each of you prefer to test or prove which isolates you have is the best performer. I was thinking of just reverting back to the good ol' PF tek to test the performance of each of my isolates to come.
How do you have it?


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12296478 -

thats how most people do it. personally start from MS and make some pf cakes. then i clone the fruit i want to agar. that way there are only a few sectors. then i usually only transfer once or twice. when i transfer i just pick the part that colonizes the quickest.


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Re: How do you (personally) prove your isolates? [Re: oh_you_know]
    #12296510 -

Ah, I see. I was thinking, because when my agar supplies arrive, I will not have any fruit bodies to use for tissue, or spores, that I will use spore solution from a syringe onto agar, and just have to make the multiple transfers, or however many it takes till I get one with no sectors. When I finally get mycelium with no sectors, I would take that and grow it out correct? Do I just make up PF tek jars, minus the verm barrier and place that agar wedge in there and allow to colonize, and fruit to check performance?


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12296552 -

i would grow it out and get it alone, then take that and make another plate. use the plate you cut and cut another piece and use half pint jars or something small so it colonizes quick and grow it out on grain. then follow this grain lc tek then do a few pf tek jars. then you got some fruits! ive yet to have any luck with liquid [spores or lc] to agar.


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Re: How do you (personally) prove your isolates? [Re: oh_you_know]
    #12296624 -

No luck with spore solution you say? Why so, whats happening with it?


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12296644 -

im not exactly sure. sometimes nothing happens. sometimes it gets some random contam which doesnt make sense. i flame sterilize and inoc in a glove box and cover hole with micropore tape and it may get a contam. but if im in the same glove box and grab a piece of grain and toss on agar, after physically opening the jar, it doesnt contam. i would stick with ms to grain then grain lc to pf tek.


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Re: How do you (personally) prove your isolates? [Re: oh_you_know]
    #12298381 -

I was thinking, since I have some MS jars colonizing right now. I will just grow out enough to get a couple fruit bodies, and I will take a print from them and use the spore print with an inoculation loop to agar. From what I have read it seems sterile procedure as far as the actual making of the print doesn't have to be too crazy, because if I get a contam I can transfer the healthy mycelium away from the contam.

So I guess that's my plan.


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12298565 -

shroomizzy said:
I was thinking, since I have some MS jars colonizing right now. I will just grow out enough to get a couple fruit bodies, and I will take a print from them and use the spore print with an inoculation loop to agar. From what I have read it seems sterile procedure as far as the actual making of the print doesn't have to be too crazy, because if I get a contam I can transfer the healthy mycelium away from the contam.

So I guess that's my plan.



A spore print >>>> agar is going to be even slower than just putting a drop of spore solution on agar.

In your situation I would just put a drop on agar, that will give you lots of sectors. Then transfer from the rhizo/fastest growth. Your next plate will still have several sectors. Transfer 2 or 3 of those to new dishes. This 3rd set of dishes should just have a couple of sectors. Transfer those again. Now you should have around 8-15 dishes with isolates, depending on how many transfers you do from each plate.

You would have to grow each of those out to find the best fruiter.

Agar transfers don't work well on pf tek, because you can't shake so it'll probably take a long time to colonize.

I have taken a syringe with water, agitated the mycelium on a petri dish, then sucked it into the syringe, and used that as liquid mycelium, which with you can inject cakes or whatever you want.


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Re: How do you (personally) prove your isolates? [Re: teesionbear]
    #12299114 -

Oh alright. Thanks for the suggestion. I was just saying using the agar to PF tek, cause I had read a post from RR who said thats what he did.

I mean basically I want to try and test the isolate as quickly (yet carefully) as possible. Using MS on rye and then cased the rye has worked quite well for me in the past. So perhaps I will transfer a wedge to rye and case the rye to see how well the isolate will perform.

So quick question. I put a drop or two of spore solution on the agar dish, and when I take the mycelium from that dish and transfer to a new agar dish, and say get some sectors out of that, and then so on transfer those to other dishes. My question is what do I do with the dishes prior to each transfer? Are they pretty much useless to me?


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12305694 -

I'm doing isolation on several strains now across two species (Ps. cubensis and cyanescens), and will be going from agar to half-pint grain jars and from there to small (really small!) trays of bulk substrate, for each of the multiple isolates expected per strain. It takes a lot of dishes...

Why? Because I want to evaluate how well they do on what I plan to grow them on, is why...:mushroomgrow::thumbup:

And then, if I get really ambitious, I'll use a potency evaluation method from the several discussed in this forum...or if not, the usual bioassay.:shrug:

Peace
-PS

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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12318224 -

Quote:
shroomizzy said:My question is what do I do with the dishes prior to each transfer? Are they pretty much useless to me?



They can be kept as backups. If you do a transfer, and it gets contaminated, you can go back to the clean original and transfer to a new dish.

If your transfers are all clean, then the older generation dishes can be used for agar transfers to grain. of course that will be multispore, but it will be a lot quicker than going from spores.

if you have isolates or multispore lc/grain going already, the old dishes can be tossed.


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12319288 -

1.)  I first dabble and figure out how I want to grow.

What I want to grow on.  And what spore 'race' I think will do well.

I don't necessarily iron out the items I might not have total control over.  But I do use a rich clean substrate that won't vary too much if I don't know what other people will be growing on.  (So a PF tek or 9er tek is fine as long as you are also doing it for other people)
You may also want to try it out on a cheap substrate like dung or wood at that point depending on what you are working with.

1a.)  Grow from multispore and mix spore 'races' if possible use a couple methods.  This may result in some interesting results and may affect what fruits and how the end fruits look and how they are genetically.

PF cakes or cup of shrooms are great.  If you mix spore races and they germinate at about the same time and if they are interspersed you can get an extremely varied result.  This will give you a great deal of various fruitbodies to play and experiment with.

2.)  Take a few clones.
Store them for reference and make a few petris for expansion.

3.)  Make spawn and grow the mushrooms.
Grow the mushrooms how you want to grow them.

4.)  Pick and keep the clones that do the best based on the grows you do.

5.)  Keep the original clone you like!!!!!

6.)  Then just do a 3rd or 4th generation and make prints so it is a bit stabilized and you can store the genes of the mushrooms you are working with.

7.)  Grow your pure clone whenever you feel like it or do some other breeding with it.

shroomizzy said:
With this being my first attempt at dabbling in the world of agar, and strain isolation's,( still waiting on all materials to arrive) just curious as to how each of you prefer to test or prove which isolates you have is the best performer. I was thinking of just reverting back to the good ol' PF tek to test the performance of each of my isolates to come.
How do you have it?


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Re: How do you (personally) prove your isolates? [Re: Raw]
    #12320515 -

The clones are not necessarily isolates until you isolate them on agar and they don't sector...

Peace
-PS


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Re: How do you (personally) prove your isolates? [Re: PrimalSoup]
    #12327810 -

I find the hardest part is potency testing. I don't trip very often and it's hard to rely on the reports of volunteers. Rarely do I get back consistent reports, even tho i ask them to dose precisely and on an empty stomach.


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Re: How do you (personally) prove your isolates? [Re: anonjon]
    #12328019 -

I was thinking the same thing about the potency. The only feature I'm gonna have a hard time knowing is best would be the potency, as I only trip once every several months.


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12331004 -

then potency shouldnt be much of an issue, should it? a cube is a cube is a cube should mean they all about about the same potency.


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Re: How do you (personally) prove your isolates? [Re: oh_you_know]
    #12331062 -

well, that's not true about potency. you get a lot of genetic variety in every multispore crop.


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Re: How do you (personally) prove your isolates? [Re: oh_you_know]
    #12331636 -

I'm sure theres an average range a cubensis can contain of the active substance. So if I can get a consistant crop where every single fruit, which is the same genetics, all contain it at the high end of the range.


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Re: How do you (personally) prove your isolates? [Re: shroomizzy]
    #12331931 -

There's no average range.  With multispore, they can range from zero to throw your dick in the dirt.  That's why we isolate.  Testing by eating them is the only way most of us will have to prove the isolates.
RR


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Re: How do you (personally) prove your isolates? [Re: RogerRabbit]
    #12332805 -

I don't agree with that one. If you use a multispore, they all have something. The specific levels vary, but in what measurement, 100th of percent?

Also testing is quite subjective. Depending on how you are feeling, the condition your body is in, your recent diet, where you are in life, and other subjective variables; people around you, setting, amount consumed, current state of the world, and so much more.

Sure you can ignore all that information and go on X weight, and write a subjective report. You could even do studies with A & B test groups. Crap I can manipulate people into thinking this batch of mushrooms are very strong and to only consume a little, and if someone has the balls to take more then I recommend, they piss in their pants. The power of influence which defines reality.


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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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