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OfflineUzimyco
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Lurker finally posting question on “inconsistency of cultivation” * 1
    #27038563 - 11/14/20 10:35 AM (3 years, 3 months ago)

Hi All

I already know that my question is vague to the point it will be hard to answer but figured I’d take a shot as the frustration is a bit more than I can tolerate.

I am a highly experienced horticulturalist in a wide array of complex genera however new to mycology.

I have had a circumstance that simply doesn’t compute to me.  On multiple occasions I have had situations where (in a truly sterile environment) I have created spore syringes, cake media, sealing process and inoculation procedures however invariably some of my jars will roar out of the gate with mycelium development. Others almost weak and unimpressive and others virtually no development at all.

Now since media is all together.  Spores are dispersed in a communal beaker prior to being up drawn by syringe and all are inoculated and incubated together, how on earth can there be so much disparity in result. Again as an experienced horticulturalist I do not understand. This is all in controlled environments and purposely the variables have been excluded as to avoid such circumstances.

Just wanted to throw it out there if there were any opinions?  It is massively frustrating to me. Note:  I have no signs of contamination in any way shape or form. Again by using protocols that are on par with laboratory settings that is not an issue I have dealt with. It is the variability of the result that I cannot fathom.

I will admit that the only thing I can think of is the “amount of solution” introduced during inoculation however I am trying to use an equivalent amount of agitated solution which may be 1/10th of a cc diffferent here or there. It doesn’t seem likely that would be enough to be so pronounced.

Oh lastly. I have had the experience with a variety of cubensis strains so it isn’t limited to one type.

Thanks for reading. Please let me know if you have any thoughts.


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OfflineCorundum
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco] * 1
    #27038592 - 11/14/20 10:54 AM (3 years, 3 months ago)

Each pair of spores that come together to form a colony is like 2 people having a kid. The genes are a random combination of the genes from the gametes. This is why multispore is so random. Consider looking into mushroom cloning techniques


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:samus::samus:


Edited by Corundum (11/14/20 10:55 AM)


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Corundum] * 1
    #27038616 - 11/14/20 11:11 AM (3 years, 3 months ago)

Because that's how fungi work. They're not seeds, they're hyphae. To increase their chances for success they have a huge genetic load that they blow containing insane amounts of DNA.

Some hyphae are great at what they do and forge forward, some suck and grow slowly.

That's why we do agar work first instead of ms syringes. Ms syringes are a crapshoot.


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27038994 - 11/14/20 03:19 PM (3 years, 3 months ago)

Many thanks for your replies.

They are genuinely appreciated. To go a step further I would very much like to take your advice and move to agar as this is wildly frustrating.

I realize that a quick search of the forums along with the internet will provide countless versions of material explaining the process. I would however much prefer to take some direction from folks like yourself as you’re experienced where I am not.

Would you be so kind as to post up a link of an agar process that is suitable however not overly complicated?  I just prefer to obtain information that has been vetted by experienced people vs stuff floating around. If you’d be willing to post a link I would be very appreciative.

Many thanks for taking the time to reply. Lurking only goes so far and I’m already glad I posted based on what you’ve provided.  I did not realize the potential benefits of agar vs a brf style tek. I can see it is obviously VERY worthwhile as going through the process for this level of hit and miss isn’t very fun.


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27039006 - 11/14/20 03:23 PM (3 years, 3 months ago)

500mL water + 7.5g agar + 7.5g LME + Pressure Cooker + 50 Petri Dishes = Agar work

Boil water, add agar and LME, blend/mix/whisk it together. Pour into jar with vent hole, cover with foil PC for 20 minutes after venting for 10. Take it out, let it cool down a bit. Move everything to SAB, pour plates.

When working in SAB use small, slow movements, don't bump the box and keep everything sterile.

Or just follow D3monic's tek, he's good at agar.

https://www.shroomery.org/forums/showflat.php/Number/26628234#26628234


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OfflineLogicaL ChaosM
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27039041 - 11/14/20 03:42 PM (3 years, 3 months ago)

What kind of spore syringe did u have? Was it mostly clear?

Ive noticed with spore syringes that have really high concentrations of spores, for example one spore print for one 12cc spore syringe, u see explosive, consistent growth as the spore concentration on the substrate is so high.

I would say spore concentration is the biggest reason for varied germination times as well as inconsistencies in the PF material (too wet, too dry, wrong ratios, etc).


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Offlinetravels_slightly
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: LogicaL Chaos] * 1
    #27039058 - 11/14/20 03:55 PM (3 years, 3 months ago)

Quote:

LogicaL Chaos said:
What kind of spore syringe did u have? Was it mostly clear?

Ive noticed with spore syringes that have really high concentrations of spores, for example one spore print for one 12cc spore syringe, u see explosive, consistent growth as the spore concentration on the substrate is so high.

I would say spore concentration is the biggest reason for varied germination times as well as inconsistencies in the PF material (too wet, too dry, wrong ratios, etc).





Wait, so by this logic it's ideal to apply as much spore as possible to induce quicker colonization? Am I missing something or is that the sum of it? I only ask because I feel like I see the same reoccurring information being shared here: smaller the amount of MS inoculate, the better. Am I mistaken?


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General supporter of all things mystifying, specially when they happen to simultaneously be incriminating. ISO spores beyond my limited collection. I make and deal in many wares, perhaps we can exchange cool goodies :smile:


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: travels_slightly] * 1
    #27039155 - 11/14/20 04:57 PM (3 years, 3 months ago)

The larger the spore density the more opportunities for mating which leads to more dikaryon which leads to more potential fruiting bodies.


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OfflineLogicaL ChaosM
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: travels_slightly]
    #27039206 - 11/14/20 05:36 PM (3 years, 3 months ago)

I think so.

My logic is that by using more spores, you are getting more genetic matches competing for each other. Its like survival of the fittest at the microscopic mycelium level.


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: LogicaL Chaos]
    #27040030 - 11/15/20 07:21 AM (3 years, 3 months ago)

Thank you for that information.

Without question the syringes were quite dilute. Not necessarily on purpose but my prior spore experience relates to rare tropical ferns. VERY different situation. Too many spore in the fern world leads to over crowding which becomes a competitive mess.

But no doubt I “could” have used much denser concentration. As a matter of fact I believe I poured like 1 cup of spore solution down the drain as I had made many syringes and figured that it was just overkill.

Again as a new person to the hobby there are some things that aren’t obvious. For example. I would think that any activity even being it a bit would just go the distance and fully colonize anything. Maybe that’s true but it would take an amazingly long time. I have some jars that have “hard core” mycelium in them but it’s the size of a quarter and doesn’t seem to keep spreading out. Don’t know why but I’m learning very fast that what seems logical to me through my experiences are highly not applicable here. This is why I am thankful for such a forum as obtaining the info is slow even when speaking with highly experienced people. I don’t want I think what it would be relying on purely books and the like.

It’s an interesting thought however would likely require a significant number of prints if you’re suggesting using a print per 10-12cc. I’ve been innoculating these half pint jars with about 3-4cc. Maybe I could barely get 3 out of 1 syringe and this process isn’t exactly overnight so it seems counterproductive unless I had like a whole bunch of prints.

I like the idea of cloning or using agar to determine the higher quality of growth “before” the process really gets underway” as it makes much more sense.

I freely admit I will have to study the agar methods repeatedly for a bit.  I know for those experienced it looks easy. I’m a bit of an expert in some areas so things as such are simple to me whereas highly complex to others. It’s a kind of understanding that develops over time. So I do need to read, re-read, re-read, etc until I really understand the whole thing.

Again many thanks to all who’ve contributed. I attest a great deal of knowledge on quite a few subjects to forums such as this.  And this one is no different.  It’s likely one of the few things that I can categorically say is “good” about the internet.

Many thanks!


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InvisibleStipe-n CapMDiscord
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27040048 - 11/15/20 07:38 AM (3 years, 3 months ago)

I would cut out the middle man and just make prints, why bother going through the trouble and resources for a syringe?
Print to foil, use inoculation loop to collect spores from foil, streak agar plate.


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Stipe-n Cap]
    #27040053 - 11/15/20 07:42 AM (3 years, 3 months ago)

:whathesaid:


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27046410 - 11/18/20 08:57 PM (3 years, 2 months ago)

Hi all. I’m convinced that I had already posted this but somehow I guess maybe I never submitted it.

Based on the information that you’ve all provided I think I have a pretty solid understanding of the agar process. Where my confusion comes in now is that after seeing the process of spawning on agar  And then further seeing the replating of quality growth on subsequent dishes, I’ve watched videos of the whole agar clump in essence dumped into a container that looks like a growing substrate such as brf? Or perhaps something else?

Is my impression correct? Once the agar spawn has been done and perhaps pieces transferred to other agar dishes to further grow out quality mycelium, is that what I am looking at? Where the entire agar disk with mycelium growth is placed into a container with some kind of nutrition such as brf to take it further?  I swear I know I posted this. I wonder if I did in in a different thread by accident?

Any of your advice again is most genuinely appreciated!


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OfflineGrinchGrower
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046467 - 11/18/20 09:18 PM (3 years, 2 months ago)

You can put the *clean* colonized agar to BRF cakes (myc slurry) OR do what most people prefer and go to bulk by dropping little pieces in sterilized grain jars/bags (spawn) then put that to a substrate (coir, hpoo etc.) once it is fully colonized.

You can also make LC (liquid culture), but this is a little more advanced.


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OfflineYeetusdeetus
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: GrinchGrower]
    #27046474 - 11/18/20 09:27 PM (3 years, 2 months ago)

Once the mycelium looks healthy on agar you would transfer a piece of it to a vessel of hydrated, sterilized whole grain such as rye, oats, wheat, barley, etc.

Brf is usually used in conjunction with inoculation methods involving liquids such as with a spore syringe, liquid culture, or liquid inoculant

Once the mycelium has colonized your sterilized grain you mix it with your hydrated bulk substrate such as coir, in a fruiting vessel (shoebox, monotub)


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Yeetusdeetus]
    #27046503 - 11/18/20 09:44 PM (3 years, 2 months ago)

Many thanks to your replies. I think that’s what I meant in essence. I will have to just find a reliable easy bulk media to use as this all should eliminate the touch And go aspect of spore syringe to brf tek.

Any recommendations as to the best formulation (general consensus)) as to the grain mix or is it simply sterilized grain as you’ve described?  I assume it is grain in its original form vs “flour” structure or at least it looks as such in videos?


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OfflineGrinchGrower
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046527 - 11/18/20 09:58 PM (3 years, 2 months ago)

The search function on this site has literal DECADES of information including grain prep TEKs, bulk substrate recipes, etc.

I suggest finding one of the Trusted Cultivator members here you think sound like they know what they are doing (just kidding, they all do) and simply following one of their TEKs step-by-step.


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Offlinetiptrippy
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046548 - 11/18/20 10:21 PM (3 years, 2 months ago)

Quote:

Uzimyco said:
Any recommendations as to the best formulation (general consensus)) as to the grain mix or is it simply sterilized grain as you’ve described?  I assume it is grain in its original form vs “flour” structure or at least it looks as such in videos?




I believe most people have been so patient and willing to help you because of how well spoken and intellectual you seem. However, that could fade very fast here for asking a bunch of questions that could very easily be answered if you did a little bit of exploring and research on the forums.

Here are a few tips:
Sticky posts at the top of the forum usually contain compiled lists of teks from many Trusted Cultivators.

Check the agar portal.
Check the Noob Forum.

Search for people like bodhisatta, pastywhyte, eatyualive, D3onic, a.k.a., verum, mushboy, mushroomnate and read through their posts and teks. You could literally spend hundreds of thousands of hours researching this website.

Finally, find a few good teks AND STICK TO THEM. Don't use a bunch of info from 20 different teks and make your own plan. That almost never works.

Good luck


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: tiptrippy]
    #27046564 - 11/18/20 10:50 PM (3 years, 2 months ago)

I can appreciate your reposnse and in fact do appreciate it.

I do know a few things:

Yes I can look on forums. I can internet search like few can. What I can tell you is I did significant reading and research and ended up deriving information that was incomplete snd misled me causing wasted time and resources.

My request was simply that all information of forums and the internet are not created equal. It is for that reason that I requested the information from you all as you were kind enough to provide valid and succinct answers to questions which have eluded me.

An example is the commentary above having to do with spore density and or spore load having a. material effect on syringe inoculation success. It is the single first thing I’ve read detailing such snd I have read more material than I care to admit.  So I urge you not to mistakenly interpret my questions as either laziness or stupidity regarding searching through the internet. Your interpretation is erroneous. It is an intellectual mind that focuses on information and the source.vs information with no inherent validity.

I don’t care to waste effort and resources on making mistakes anymore. That is what has occurred and that is what prompted my post. All is not what it appears on the surface.

Again however I can appreciate your feedback.


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InvisibleStipe-n CapMDiscord
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046568 - 11/18/20 10:58 PM (3 years, 2 months ago)

You seem like a thoughtful, intelligent human. Stop making spore syringes, they're for noobs.

This is how to succeed:

1. Acquire spore print; (if you have a spore syringe just Streak the solution)

2. Streak spores to agar using an inoculation loop;

3. Clean culture through isolation;

4. Use clean agar to inoculate grains or liquid culture;

5. Use grains to spawn tubs, or use as masters for gtg. If you made LC with your agar then use it to inoculate the grains that will be used for tubs.

Fuck spore syringes, fuckin forget that they even exist. Spore syringes can eat a dick....also, spores to grains or LC is fucking retarded, don't do that. If you become a mushroom growing God and feel like fucking around with any of that shit after you've learned how not to fuck everything up, feel free....until then, fuck all of that.

Any issues that you've previously experienced are directly correlated to your failure to follow that protocol. 

This is the way.


Edited by Stipe-n Cap (11/18/20 11:38 PM)


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