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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19036264 - 10/26/13 03:45 PM (10 years, 3 months ago) |
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1) I still wonder if your prints are still actually sterile. I'm not saying "they're not".
Pins are very aggressive when transferred and will usually outrun most contams, so I'm not sure that's a perfect qualifier.
Even if they are not totally sterile, I bet they are at least clean as hell and worth the effort.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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MaJiK_420
...lost



Registered: 06/30/08
Posts: 447
Last seen: 6 years, 1 day
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: SpitballJedi]
#19039218 - 10/27/13 03:02 AM (10 years, 3 months ago) |
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You sure do catch a lot of flak on your posts Violet. You go against the grain and think for yourself, and I appreciate that. I still love doing monotubs, who doesn't love seeing a tub full of mushrooms? but you got me hooked on the plastic containers and I like some of the options you present here.
I wish I had a picture... actually I think I do somewhere, but im too tired to find it, but I had a colonized pp5 that I emptied out for spawn and then put a lid on it and it went in my dirty bin. finally get around to cleaning a week or two later and just from the seeds stuck to the side of the plastic, they all networked together and grew a mushroom from bottom to top and halfway back down. I dropped my jaw, is was spindly and the cap was tiny, but it had to have been 6-7 inches. Anyway I thought it was pretty remarkable. Amazing that such little can still produce fruit, like your rice patties. good stuff.
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: MaJiK_420]
#19040342 - 10/27/13 11:32 AM (10 years, 3 months ago) |
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Quote:
MaJiK_420 said: You go against the grain
actually its WITH the grain...... but ,i see your point lol.
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: anne halonium]
#19040614 - 10/27/13 12:37 PM (10 years, 3 months ago) |
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I'm pleasantly surprised. 
Some of these ideas will make it into my future grows.
I've been on the pp5 train since I started, it's an obvious vessel for this hobby compared to glass.
As for the grain water/ malt extract being substitutes for eachother. Wouldn't the mushrooms have the enzymes necessary anyway to treat both as the "same" 150F mash turns starches to sugars by enzymatic reaction so LME has sugars more than starches that the grain water would have. Either way the mycellium can eat it and turn it into the same amount of energy I would imagine.
Edited by Trusted cuItivator (10/27/13 12:43 PM)
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OgreLokon
Pretty Fun Guy


Registered: 06/30/13
Posts: 512
Last seen: 6 years, 5 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: bodhisatta]
#19041148 - 10/27/13 02:39 PM (10 years, 3 months ago) |
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I love reading your Teks, Violet! Always detailed and very well written.
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MaJiK_420
...lost



Registered: 06/30/08
Posts: 447
Last seen: 6 years, 1 day
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: OgreLokon]
#19042073 - 10/27/13 06:46 PM (10 years, 3 months ago) |
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I'm gonna try the brown rice thing, but I was wondering, how do you prepare the brown rice?
I assume you hydrate it and let it dry... but any specifics? I'll do a search but I figured it might be something worth addressing here.
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: MaJiK_420]
#19042120 - 10/27/13 06:55 PM (10 years, 3 months ago) |
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Quote:
MaJiK_420 said: I had a colonized pp5 that I emptied out for spawn and then put a lid on it and it went in my dirty bin. finally get around to cleaning a week or two later and just from the seeds stuck to the side of the plastic, they all networked together and grew a mushroom from bottom to top and halfway back down. I dropped my jaw, is was spindly and the cap was tiny, but it had to have been 6-7 inches. Anyway I thought it was pretty remarkable. Amazing that such little can still produce fruit, like your rice patties.
It absolutely is wonderful. A single grain can sustain a mushroom 10 times its size, as long as the network can pull moisture from anywhere. Quote:
anne halonium said:
Quote:
MaJiK_420 said: You go against the grain
actually its WITH the grain...... but ,i see your point lol.
I love a good pun! Lamer, better!
Quote:
MaJiK_420 said: I'm gonna try the brown rice thing, but I was wondering, how do you prepare the brown rice? I assume you hydrate it and let it dry... but any specifics? I'll do a search but I figured it might be something worth addressing here.
And someone got iffy with me about posting my grain preparation... Rice is easy to prepare, just have to make sure you don't overdo it!
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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MaJiK_420
...lost



Registered: 06/30/08
Posts: 447
Last seen: 6 years, 1 day
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19042157 - 10/27/13 07:02 PM (10 years, 3 months ago) |
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Awesome, thanks for the quick reply.
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wildernessjunkie
Reshitivest



Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: MaJiK_420]
#19050205 - 10/29/13 02:53 AM (10 years, 3 months ago) |
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19052520 - 10/29/13 03:40 PM (10 years, 3 months ago) |
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Quote:
Violet said:A single grain can sustain a mushroom 10 times its size
You really are redefining science. Is that by weight or by size? Need clarification before I mathematically disprove the possibility of this ever happening.
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: barong]
#19052725 - 10/29/13 04:22 PM (10 years, 3 months ago) |
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Quote:
barong said:
Quote:
Violet said:A single grain can sustain a mushroom 10 times its size
You really are redefining science. Is that by weight or by size? Need clarification before I mathematically disprove the possibility of this ever happening.
Dry a mushroom and grind it into a powder, you'll see just how little non-water material a mushroom is. I do doubt that a single grain can make a mushroom 10 times its size, but a single grain can make a mushroom large enough you wouldn't imagine it having came from a single grain.
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: bodhisatta]
#19052968 - 10/29/13 05:09 PM (10 years, 3 months ago) |
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Quote:
bodhisatta said:
Quote:
barong said:
Quote:
Violet said:A single grain can sustain a mushroom 10 times its size
You really are redefining science. Is that by weight or by size? Need clarification before I mathematically disprove the possibility of this ever happening.
Dry a mushroom and grind it into a powder, you'll see just how little non-water material a mushroom is. I do doubt that a single grain can make a mushroom 10 times its size, but a single grain can make a mushroom large enough you wouldn't imagine it having came from a single grain.
For weight, it's not even close to being possible. Grain vs hydrated grain is a ratio of roughly 1:16. For a mushroom that's 90% water, how on earth is the transfer possible? It defies logic and completely goes against bioefficiency observations from RENOWNED AND CREDIBLE GROWERS such as Paul Stamets & Marc Keith
Let's look at it another way. For every 3 grams of grain, would you expect to get 30 grams of wet shrooms? If so, then this TEK truly is magical, to turn a tiny few grains into a full trip.
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: barong]
#19053270 - 10/29/13 05:59 PM (10 years, 3 months ago) |
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For every 3 grams of grain, would you expect to get 30 grams of wet shrooms?
Your getting confused with mass and density. 3grams of grains wouldn't produce 30 grams of wet shrooms for sure, but 1 grain might weigh 1g make a mushroom that's 3g wet but looks a hell of a lot bigger than the single grain.
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: bodhisatta]
#19053989 - 10/29/13 08:01 PM (10 years, 3 months ago) |
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So the question remains. Where's the clarification on "TEN TIMES" a single grain? Whether by volume or by weight, it's a ridiculous statement
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: barong]
#19057234 - 10/30/13 11:59 AM (10 years, 2 months ago) |
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First you play as if a very general statement about grain nutrients in agar is saying something entirely different about malt, and now that little statement is "redefining science"? Can I not say something in an unscientific fashion without some anal-retentive person pretending that I'm trying to pass it off as a scientific certitude?
For starters, I did not say the grain is alone. You assumed my statement literally means that you can accomplish such a grow on nothing but a single grain or by proportion. That sure would be an easy thing to fail, yes? Why didn't you think I said a single grain can physically support a mushroom ten times its size? Because, well, that's mainly what I meant.
Even further: Many of the fruits I grow recently grow from absolutely nothing except their own network on the side of plastic. As long as they can bring what they need from anywhere, typically less than 5 inches, fruits can from anyhow.
It's quite clear that I wasn't making a yield claim of some sort. I do talk about yield ratios and that statement not what it looks like. I'm always honest about my results, why would I lie?
But to make a bit of a point (and oh how I do love it when detractors inadvertently set me up for one) lets indeed approach the question mathematically as you say, and see how unduly your incredulousness is:
I'm sure you understand how B.E. is calculated? Since I grow straight-grain my isolate cultures easily reach 300%+. The grain petris in this thread have around 22g of rice in each. Solid isolates give me up to 7-8 grams from these little cakes, a BE of 320-370%
Since mushrooms are ±90% water, and 1mL (1 cubic cm) water weights 1g, it's adequate to say that a 10g mushroom contains 9mL water and will be over 9 cubic centimeters.
With a BE of 350% from straight grains via bottom-watering, by proportion 10g of rice can grow 35 fresh grams of mushrooms, ~35 cubic centimeters.
This is what 10g of rice looks like:
 
This photo is the closest I have to 35g fresh:

About 3 times the volume, and even looks like more.
So even if I were saying your spin on my statements, at worst I would be exaggerating some. At best you would be focusing on a trivial point, removed of its meaningful context, in an unsuited light. Besides, you may not fully understand science in this regard: Science doesn't "disprove" anything, it only succeeds or does not succeed in proving a hypothesis. Edgy scientificky-advocate-type people just think science disproves things they don't believe in.
I have no doubt in your ability to "disprove" something of that sort - all you have to do is fail the grow; anyone can do that and think he/she has "disproved" something, especially if what they think they disproved wasn't even the real hypothesis so-to-speak. What counts is how far one can succeed! With ideal process and great culturing, which is what this thread is about, the wonderful results I actually do lay claim to are not difficult to reach!
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing Tech • Agar containers, [Re: Violet] 2
#19057440 - 10/30/13 12:40 PM (10 years, 2 months ago) |
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i think they miss the larger point violet, and, mainly seem to parse statements to be annoying on details.
i find it odd, how the bucketeers, only apply scientific reasoning to grain grows.
im not sure i would go for 10-1 ratio, but, i know from experience, the sub weight ratio from bulks , is off the charts lame , compared to grains. of course, parsing words, makes peeps overlook that point.
most of the bulkers, dont do bulks because they love feces. they do bulks because they dont have skills for grains, and alot of em, havent done grains to scale, so they really dont know any better.
when someone shows decisively, that grains yield more, bucketeers are left with the choice of limited teks, or actually cleaning their labs and upping the skill. ( or, be outdated) so,the rational response? parse words, attack threads on minor points.
to admit grains have advantage, when used with skill, is NOT an option, for some of them.
1-1 or 1000-1 , minor point. big point is this "grains kick ass 100% of the time."
buckeeters, would never guess on their ratio, as no matter what it was, it would invite the obvious comparisons.
peeps out front , do things all the time, that cause great debate on the most minor of details. after while , the most geeky scientists , actually measure such stuff. by then, no one cares, cuz , it either performs, or it doesnt.
i suppose , some peeps need a data sheet, and some dont ,to see the obvious.
Edited by anne halonium (10/30/13 01:02 PM)
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jpack666


Registered: 10/01/13
Posts: 484
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19075474 - 11/02/13 02:41 PM (10 years, 2 months ago) |
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Hi violet, been reading your post after seeing the link in mine about seringe vs print if transfering to agar. Nice tek!
But as I was reading it, I kept asking myself what experiences led you to get involved in these extra steps... I suppose being fed up of contaminants in agar from unclean prints?
Maybe something else....
If I buy a print online, is it gona require isolating the culture from the contams (in petri dish) with a couple of petri transfers?
Or is this more of an issue when we make our own prints because we don't use the same environment or have te same goals or budget as these online suppliers?
So if prints are not sterile, does it mean you are garanteed not to have a pure culture on your first agar plate?
How many isolations/transfers do people usually go through to get pure culture? (Not lab conditions, more like home project conditions with glove box)
-------------------- "There's a negative and a positive to everything." For more information, RogerRabbit's Website/Videos
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: jpack666]
#19075646 - 11/02/13 03:21 PM (10 years, 2 months ago) |
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Quote:
jpack666 said: as I was reading it, I kept asking myself what experiences led you to get involved in these extra steps... I suppose being fed up of contaminants in agar from unclean prints?
What extra steps?
I'm sure you'll understand a bit better once you have some grows under your belt. These aren't extra steps at all - in fact some of them skip several steps from other grow procedures.
Taking a clone is only possible after a fruiting grow begins pinning. This technique allows one to get to the clone point with a single dish and makes the sterile work of taking a clone far easier. It skips all the steps of a full grow-out and eliminates cultural hyper-expansion, leaving the culture far more young than it would otherwise.
In fact the whole tek is about skipped steps and shortcuts. No petris regularly ordered online, no parafilm clingwrap or bags, no extra materials for agar, no careful pour timing or procedure, few transfer tediums, nearly no isolation from contaminants, no dud cultures, no large grows of chance with multi-spore, no fruiting chamber, no automation... all the while producing a bonus yield and resulting in young, strong, ideal cultures!
Quote:
jpack666 said: If I buy a print online, is it gona require isolating the culture from the contams (in petri dish) with a couple of petri transfers?
Hopefully you'll have bought from a place with good clean standards for their product. You are quite unlikely to have contaminations from such. Wild-card vendors or trades from growers of questionable quality might cause some real clean-up work though, or even total failure...
Quote:
jpack666 said: Or is this more of an issue when we make our own prints because we don't use the same environment or have te same goals or budget as these online suppliers?
If you take a print from a mushroom grown in gnarly environs there is a variable chance of issues. If you take them from sterile invitro, you will have NO issues, as long as your clean tech is good.
Quote:
jpack666 said: So if prints are not sterile, does it mean you are guaranteed not to have a pure culture on your first agar plate?
Depending on the environment, and how soon the cap was picked, and how clean of a printing method was used, it's possible either to require lots of transfers or to have a totally clean dish right-off. When it's so easy to take invitro prints I don't see a point in gambling.
Quote:
jpack666 said: How many isolations/transfers do people usually go through to get pure culture? (Not lab conditions, more like home project conditions with glove box)
This is kindof like asking how many beans are in the jar. If the contaminant isn't wide-spread, and the transfer is done well, it may only take that one transfer or one more. This is usually my experience. Perhaps someone only semi-experienced with still-air box work with a very contaminated petri could end up needing lots of plates... poor souls.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19140896 - 11/15/13 01:01 PM (10 years, 2 months ago) |
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May it suffice to say that it's best to transfer pins as early as possible into maturity.
Some sterile pins transferred quite late:
    At a certain point they may even continue to mature and open caps. We definitely don't want that for a clone dish.
These dishes were all started with small pins:
      Growth is much more ideal; these cultures are and will continue to be more healthy and vigorous.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
Edited by Violet (11/15/13 01:13 PM)
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twistedty
Forcefully Retired



Registered: 07/01/12
Posts: 5,487
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19140939 - 11/15/13 01:13 PM (10 years, 2 months ago) |
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you freeze grain water?
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