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Developing Methods

Where's all the science at?

experimental research

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#21 CatsAndBats

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Posted 02 February 2015 - 07:48 PM

Technically the PC should get rid of everything alive/potentially alive.

 

Regarding the microwave, anything at field capacity should be sterilized due to the electrons in either fats or water (in our case) being accelerated enough to generate heat/energy. I am doing @wharfrat 's coco coir/coffee recipe with zombie tubs sterilized in the microwave and then noc's with colonized grain..



#22 coorsmikey

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Posted 02 February 2015 - 07:53 PM

I seem to have misled many people when I say open air. I work in a still air room. A small closet. I follow sterile methods buy using lysol, rubbing alcohol, gloves, torch lighter, take a shower before work and brush teeth. I have just now start implementing another way to help curve contams and that is to take a box and place it on the side and then work in the box. I also take advantage of self healing pots in my lids but I did just run a small test a few weeks ago to see the contam ratio of no sterile tek besides steaming some WBS jars for 8 hours. I started out with 5 jars. I did not use ANY sterile method when inoculating and did not work in my still air room. 3 jars were inoculated with MS and 2 jars with a pin clone (whole mushroom, non-sterile). I ended up losing 1 MS jar and 1 clone jar to trich. The other 3 colonized healthy. So, thats a 60% success rate or a 40% contam rate with any sterile method just to see what would happen. 
 
 
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Now, when following my preferred teks for growing bulk my success rate is nearly 100% when working in a still air room. What I do is make several master grain jars then suck up the mycelium out of them for an instant LC/LI (not sure what we consider this). I then start generating more spawn by knocking up grain bags with the liquid mycelium. Following by spawning to a sub of coir/verm. I used to use the bucket tek for pasteurizing my sub but recently switched to oven pasteurizing. Usually spawn close to a 1:1 ratio of spawn to bulk. I figure 1 dry oz of cubes per quart of spawn is a good indication of clean spawn to keep the qualitative comments at by since my definition of clean spawn is quantitative.  
 
 
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My goal is to be able to eliminate the need for people who can secure a clean spore syringe of a PC for bulk. I know that their are several methods already that accomplish this goal but I want to find a way with less work and risk. This will take time as any good experiment would.  I'm still running several preliminary projects to get a feel and idea on what to look more into and what has a good chance of working. If you notice in the mad scientist sub-forum that I have several tubs that are intended to watch how cube mycelium responds to different stimuli. I have been watching my mycelium run. lol I'm still researching for more information on possible ideas and am current reading a few micro-biology books to get a better understanding how the bigger picture. When I can get some constant results of the goal I am after I will share with the community. As of right now though, I dont have much to share.  :wub:

I am interested in seeing the tek that requires no PC for the prep of grain and agar that is less work and less risk, as far as doing bulks I have never used a PC to prep my bulk substrate, I do know people will us a PC sometimes just because of the capacity it holds. For many a PC is the largest container the have on hand for a pasteurization. There are a lot of us out there that do open air transfers and shitty pasteurizations jobs with great success. Over the years the culture of this community has evolved and we tend think how posting such is not that good for the newb just starting out. We also care about how we are perceived by the outside scientific community to a certain degree. So that is why you don't see a lot of teks that don't require sterile techniques. I don't know your whole situation or exactly what you have planned. I look forward to see what you have to offer, I just hope you consider that we have been getting away with doing stuff without a PC for a long time and as a collective we believe that learning sterile technique is probably the the best most efficient way for someone to learn, they will eventually develop their own bad habits after that. And hopefully when they learn those bad habits they can get away with, that they continue to teach others the importance of sterile technique.
But hey? Am I any better? No! I am working on an additive that creates primordia formation. What type of bad habits has brought me here.
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#23 CatsAndBats

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Posted 02 February 2015 - 07:59 PM

Regarding the above ^^^ that's why we have this forum and the mad scientists forum.. Technically @blackdustt and myself still qualify as "mad" (as hatters of course)..


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#24 coorsmikey

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Posted 02 February 2015 - 08:46 PM

Thanks for your understanding Cat!
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#25 happy4nic8r

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Posted 03 February 2015 - 01:33 PM

Without experimentation there wouldn't be most of the things we enjoy in our lives. 

 

Thinking outside the box, and trying new things, especially with using what you "have on hand", is totally withing the scientific realm if it is well documented and can be repeated.

 

There is a tendency to think just because someone is new at this, that they need to follow the tried and true.

 

Depending on the person, it might make more sense to start with a kit.

 

I have never heard of anyone saying that's the way they will always do it.

 

They buy ONE kit, complain and start their own process.

 

That's progress.


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#26 mydarling

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Posted 05 February 2015 - 11:19 PM

Nice happy4nic8r -- I like that attitude!

blackdustt -- Sounds like you're working on some interesting things, would love to see it all laid out when you get around to it.
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#27 happy4nic8r

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Posted 09 February 2015 - 02:46 PM

I had a jar of spore water, that was empty almost after filling syringes.

 

I put some tap water in it and added some Karo syrup and it started growing wispy white (I don't want to describe it like it looks but...) and it's spreading out in kind of a clump in the solution.

 

Could this be spores germinating and creating a liquid culture?

 

I have not done this before, or seen the recipe for this by chance, I have not really searched, but this seemed like a good topic header for new stuff.

 

This is new to me.



#28 Juthro

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Posted 09 February 2015 - 03:16 PM

I would say you definitely have a liquid culture, 4nic8r. But one of the draw backs of LC is, until you test it on something by growing it out, it is almost imposable to tell what is actually growing in your soup.


If it were me I would grow it out some and then try a couple of BRF jars for a sample and see if they grow clean.

Edited by Juthro, 09 February 2015 - 03:16 PM.

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#29 happy4nic8r

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Posted 09 February 2015 - 04:26 PM

Thank you Juthro. 

 

I also can try it on agar, I suppose, 1 drop?

 

See where that goes.

 

I would think if it was a perfect world, that this lc would save some time doing brf jars, or any other spore to ?



#30 Juthro

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Posted 09 February 2015 - 05:39 PM

Yes, LC is faster the muti spore. You don't have to wait for the germination process.

It is also a great way to get a massive amount of yield out of a minimum amount of product.

One or two drops of spore solution can be coaxed into a large jar of LC, a project multiplier if you will.
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#31 happy4nic8r

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Posted 09 February 2015 - 05:58 PM

I'm glad I found this out.  Not very widely advertised, at least not the recipe for making liquid cultures.

 

I've heard it mentioned, but didn't really know what that was.

 

Now I have some.....yeee!!



#32 roscoe

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Posted 09 February 2015 - 06:40 PM

 

 Not very widely advertised, at least not the recipe for making liquid cultures.

 

Uhhhh.. . . There is a whole forum dedicated to nothing but liquid cultures.

 

https://mycotopia.ne...neydextrose-qa/

 

Found in the vaults?

 

https://mycotopia.ne...the-new-vaults/


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#33 Juthro

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Posted 09 February 2015 - 07:08 PM

Thank you Roscoe, I was just about to look up and post that link to the vaults. The volume of info in the vaults is staggering, every one should look though once in a while.

It seems (to me anyway) lately a lot of folks have gravitated away from corn syrup style LC's, and are working at "milking" mycelium, or at making a slurry.

As in injecting sterile water into a healthy colonized grain jar and shaking the hell out of it and then sucking the suspended myc/water up with a sterile syringe. If you work through a SHIP, this is a easy way to keep your projects clean. As it is live myc and does not need to germinate, it is notably faster then muti spore.

The big advantage to doing it like this over an LC is that you are drawing from a known healthy culture, so it is statistically more reliable if you follow good procedure.
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#34 CatsAndBats

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Posted 09 February 2015 - 07:18 PM

IME one can store in the fridge for a while.
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#35 hyphaenation

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Posted 09 February 2015 - 08:54 PM

Milk needn't be kept in the fridge so long as you keep it in a cool place below 50 degrees... I keep mine in a box in my cold basement (Canuck winter) but even in summer its 50-ish F or below. Put it in the fridge if you want (never the freezer)but its not mandatory. No need for sugars in stored milk. I've used more than 1 year old stored mycelium water with no issues. Soon I'm going to try a two year old batch that looks good still.
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#36 hyphaenation

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Posted 09 February 2015 - 09:12 PM

 

 

 

 Not very widely advertised, at least not the recipe for making liquid cultures.

 

Uhhhh.. . . There is a whole forum dedicated to nothing but liquid cultures.

 

https://mycotopia.ne...neydextrose-qa/

 

Found in the vaults?

 

https://mycotopia.ne...the-new-vaults/

 

^^^ Bingo!

 

Perhaps you may have heard of our archives ? 

 

Liquid Culture: Karo/Honey/Dextrose Q...

http://mycotopia.net...html?1108528324

 

It's the backbone of this site ... 

 

A little reading goes a long way around these parts.


Edited by hyphaenation, 09 February 2015 - 09:13 PM.

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#37 happy4nic8r

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Posted 09 February 2015 - 10:05 PM

I see a lot of old material, not bad but a lot of reading, and a lot of it to do with injection ports, etc.

 

maybe it's time for an update on this method.

 

Let's not have all my hours of reading go to waste.....



#38 mydarling

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Posted 09 February 2015 - 10:07 PM

Ok, so no experiments but questions on LC's/slurries/milks ... alright, are we getting somewhere? :P
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#39 Juthro

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Posted 09 February 2015 - 10:17 PM

That "old material" is EVERYTHING brother. If you cant learn it somewhere in there, you cant learn it.

You think you have new idea's and thoughts, but I would be willing to bet most, if not all of them are in the vaults.

Don't underrate them when you haven't but just skimmed the topics.


Edit: Sorry Mydarling, I didn't intend to derail your thread. I get distracted easily.

My apologies if I sent it (your thread) down a wormhole.

Edited by Juthro, 10 February 2015 - 12:09 AM.

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#40 hyphaenation

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Posted 10 February 2015 - 02:42 AM

Mydarling... just an idea, but why not teach by example?... where is your experimental scientific process thread/grow?  Can you link us to one? To be honest, and no offense but in all my years hear I don't quite recall you doing anything along the lines of what your talking about here. Don't get me wrong , I've seen some killer grows by you and I agree with you it would be beneficial and good. But where is YOUR input, your experimental grow?

 

... Again, I mean no offence. In my opinion those kinds of grows will come in due course, organically, but you can only encourage, you can't force with a side of attitude (you seem a bit pissy about it).  Lets see what YOU got if you want scientific/experimental grows so bad. If you were posting said grows yourself and saying "come on everybody, join me" that would be different. Do you get where i'm coming from? I'm guessing not but I want to give the benefit of the doubt.

 

It was hard to write the above because I respect you , but I can't quite understand the anger behind your wanting these new "scientific process" rigorous grow threads. And I definitely don't understand why you don't take the bull by the horns and write up a thread yourself with new ideas. Could you enlighten me? Please ...


Edited by hyphaenation, 10 February 2015 - 02:54 AM.

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