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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: twistedty]
#19141848 - 11/15/13 05:37 PM (10 years, 2 months ago) |
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Awesome post Violet! just bookmarked it as well as ur grain prep. gotta try this, cause I hate throwing away plastic petris, and I had no idea agar could be done so well with the PP5's. 1. if u do get a contam, do u just pc the plastic jar before using it again? 2. going along with twisted's Q above, u freeze the grain water for later use, then when u r ready, take it out, defrost, and then.....add agar and pour then PC? I got lost a bit on the initial post about ur media prep I guess. U employ a no pour style, right? 3. the PP5's of agar, they are not completely sealed in PC? just a tiny twist less that tight? 4. I know it was asked before above, but to clarify, when fruiting the grain containers, the FAE is provided by the loosening the lid after 100%? 5. And ur idea behind it (one of ur many ideas in the post) was that this container of fruits, with just a loosened lid, would be a much better place to take pins from to clone because of the relatively low chance of con tams entering fruiting area vs. SGFC, Mono spawned in open air, etc. correct?
Sorry for all the Q's there are so many good ideas here! gotta try this, but gotta come correct!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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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: blindingleaf]
#19144469 - 11/16/13 07:53 AM (10 years, 2 months ago) |
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Quote:
blindingleaf said: 1. if u do get a contam, do u just pc the plastic jar before using it again?
Personally I don't. Kindof a waste of a PC run. I can understand why others might do this, but I already have a great contam control due to my tek etc., and it could only do any good with mold contams anyway which I almost never see. I just dump out the contam'd container in an appropriate place outside, and bring it right back in to the sink to be washed out.
Quote:
blindingleaf said: 2. going along with twisted's Q above, u freeze the grain water for later use, then when u r ready, take it out, defrost, and then.....add agar and pour then PC?
I freeze it when it will be used MUCH later, like the pulpy stuff I have for future agar grows (not culturing). Usually the grainwater for agar dishes doesn't get frozen as I'll go thru it in a week or so; fridged rather.
You can add agar-agar to the grainwater then distribute, but you have to make sure the agar mix is even before every pour, as it will settle especially if the liquid is cold. Different strokes for different folks, but I just weigh out around .5g agar-agar per container with 20-25mL grainwater already loaded in them. This ensures the agar is distributed evenly, avoiding leaving some of them too soft and others too hard.
Quote:
blindingleaf said: 3. the PP5's of agar, they are not completely sealed in PC? just a tiny twist less that tight?
That's right! Although a 'tiny' twist may be too little depending on how tiny you mean. 1 inch turn is great. Between 1/2" and 2" is the loose range without falling off.
The containers all say "UNSCREW LID BEFORE MICROWAVING" – that's pretty much exactly what we're doing here, so it could be said it's simply following the container's directions!
Quote:
blindingleaf said: 4. I know it was asked before above, but to clarify, when fruiting the grain containers, the FAE is provided by the loosening the lid after 100%?
Yes, when invitro fruiting grain containers, as shown in this thread
Quote:
blindingleaf said: 5. And ur idea behind it (one of ur many ideas in the post) was that this container of fruits, with just a loosened lid, would be a much better place to take pins from to clone because of the relatively low chance of con tams entering fruiting area vs. SGFC, Mono spawned in open air, etc. correct?
Yes correct. Not just a "low" chance of contams, but nearly NO chance. I ended up taking at least 24 pins in this last culturing round and had not a single contaminant. Not only does it have a great sterile success chance, but doesn't require splitting and/or cutting a mature mushroom for a tissue sample, ending up with a clean and vigorous culture with less work and expansion, needing only sterilized tweezers to pluck and drop a pin. All of my cultures have been isolated with just 2-4 transfers!
-------------------- 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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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19145797 - 11/16/13 03:32 PM (10 years, 2 months ago) |
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awesome! thx for the articulate response! I love this idea, super creative thinking outside the box. Hopefully thread keeps going, cause I wanna try this and post pics after my current project! do u PC ur tweezers or just flame?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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dr.alkaline



Registered: 12/15/12
Posts: 684
Last seen: 4 years, 9 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: blindingleaf]
#19152112 - 11/17/13 10:01 PM (10 years, 2 months ago) |
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For a 30oz twist top container (Ziplock twist-n-loc brand medium) filled 1/4 way up with grains, about how much water do you recommend adding as "bottom water". I know you mentioned adding more bottom water after the first flush, but is this done in between each and every flush?
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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: dr.alkaline]
#19153146 - 11/18/13 08:13 AM (10 years, 2 months ago) |
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Doesn't take much, we can only add a bit of water at a time. 1/2 inch to 1 inch is fine, no more than halfway up the cake is my suggestion
Not only is it done for every flush, but full-fruiting genetics will need it at least 2 times per flush to grow what they're capable of!
In fact certain grains don't hold enough water for what they could put out in the first flush. With some grains (particularly RGS) I get 2nd to 3rd flushes the same size as the 1st since the water starts flowing after the 1st flush. Rice seems to hold more water and ends up calling for less bottom-water. So it all depends on the status of each cake.
-------------------- 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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Mosey3012



Registered: 12/12/12
Posts: 675
Last seen: 3 months, 15 days
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19153870 - 11/18/13 12:24 PM (10 years, 2 months ago) |
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Well so far I've had 0 luck with using the RGS soak water mixed with agar. As instructed I've added .5g agar per 25ml "soak water" and PC'd like I would my normal (MEA) agar recipe. As a test I've tried a few different things, the final test will be a fully colonized grain plucked from a jar and placed on it prior to a G2G.
I've attempted 2 separate spore germination's (1 by simply scraping spores from the print to fall onto the agar, another using an inoculation loop which I would dip into the agar prior to rubbing it on the print then swooshing it around on the agar itself) - 2 weeks have passed and nothing (those same spores have germinated on rye berries already)...
I've also attempted 3 separate clones along side my regular agar recipe and none of those piece have done anything other then sit there while my regular agar (MEA) recipe has shown signs of regrowth.
Not sure y this would be, made quite a few of these plates whenever I read this tek so perhaps the mixture I used was 2 rich in nutrients or something? As stated above my final test with this particular mixture will be to see if a fully colonized grain will continue its growth out onto the media. Will be reporting back then.
Plan on trying the rice water next and another version of my malt mixture as it seems that not matter what I do with my current mixtures spores just wont germinate to save my life. Im assuming this means the mixture is 2 rich for the spores...
Note- the MEA recipe im using is (6g of malt and 5g of agar to 340mL water) - would that create a mixture 2 rich for spores?
-------------------- "Each of these lives is the right one! Every path is the right path. Everything could have been anything else and it would have just as much meaning"
  "All that is gold does not glitter, Not all those who wander are lost"
Edited by Mosey3012 (11/18/13 12:32 PM)
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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: Mosey3012]
#19154688 - 11/18/13 03:15 PM (10 years, 2 months ago) |
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That's quite strange. Haven't heard of that happening before. The cause is surely not "too much nutrition", I've grown full flushes on much greater concentrations of grain nutrient suspensions.
When you say "soak water", if you mean that the solution you're using is just water that grains were literally soaked in, then it's hardly grainwater at all. It takes a boiling prep to really get grainmatter suspended in the water.
One thing is certain: Grainwater agar works. Your problem isn't due to the grains themselves. Maybe a fungicidal seed coating, in which case food-grade rice will work for you as well as untreated seed. Maybe it's something else...
Keep trying stuff! Isolate your variables. For instance, if myc grows on the whole prepared sterile grains but not grainwater agar from those very grains, your problem is something you're adding to the grainwater after prep, or the likes.
-------------------- 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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RogerRabbit
Bans for Pleasure



Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 4 days
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Re: Culturing • STERILE SPORES [Re: Violet]
#19154749 - 11/18/13 03:24 PM (10 years, 2 months ago) |
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Quote:
Violet said: STERILE SPORES Considered an oxymoron by most that I've seen on these forums. It's often said that "No spore print is sterile, since fruits do not grow in sterile conditions." These ones do.
Sterile spores will not grow. I think you meant to say 'clean spore print' or something similar, but not sterile. The term sterile spores is only an oxymoron if you expect them to grow. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing • STERILE SPORES [Re: RogerRabbit]
#19155025 - 11/18/13 04:06 PM (10 years, 2 months ago) |
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Um, that was already brought up...
Quote:
Mush4Brains said: If spore prints were sterile nothing would grow from them.
Quote:
SpitballJedi said: In this context, "sterile" means does not contain anything but the desired spores.
I say "sterile spores" in the same way that we do "sterile work" despite that we are actually breaking sterility by inoculating the target organism. If we did "sterile work" and "kept our jars sterile" as strictly as you imply "sterility" to mean there, we'd never grow anything in our jars either...
But perhaps it would be good to change the headline to "sterile spore printing" to stop drawing posts for such a thing
-------------------- 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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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Re: Culturing • STERILE SPORES [Re: Violet]
#19155230 - 11/18/13 04:37 PM (10 years, 2 months ago) |
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After reading this thread, there is a huge amount of useful and mind-expanding information, coupled with a ton of shit, and even detrimental information.
As a whole, I'd say the good outweigh the bad.
I will NOT go deeper into this statement, I do not plan on starting an argument on this thread, though I feel I am able to voice this simple opinion.
My second opinion, Is that anne halonium and violet..
are clearly the same person, or have a seriously close relationship, even cultish.
Adios
--------------------
The ego is nothing other than the focus of conscious attention.
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Mosey3012



Registered: 12/12/12
Posts: 675
Last seen: 3 months, 15 days
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Re: Culturing • STERILE SPORES [Re: Violet]
#19155311 - 11/18/13 04:48 PM (10 years, 2 months ago) |
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The soak water im referring to was the left over water from my tote after "flash hydrating" the seeds as you described doing before changing the tek up into parts whenever you wrote out these newer ones you just did. The exact same redish water stuff u have pictured right below "grainwater agar" in this tek to the right. I simply jarred that up after the grains were removed then a few days later added .5g agar to every 25mL plate I planned on using. Heated it up a bit on the stove and poured them, then into the PC as usual. I totally get what you're saying though, thus my posting about it. Was hoping to see where in the process I goofed.
EDIT: Went ahead and moved off topic info onto correct thread
-------------------- "Each of these lives is the right one! Every path is the right path. Everything could have been anything else and it would have just as much meaning"
  "All that is gold does not glitter, Not all those who wander are lost"
Edited by Mosey3012 (11/18/13 05:01 PM)
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing • STERILE SPORES [Re: JMcDoogle] 1
#19156216 - 11/18/13 06:58 PM (10 years, 2 months ago) |
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Quote:
JMcDoogle said: even cultish.
glad to see ya on board " J", even my detractors are a cult.
good teks inspire the grow. violet has some good teks. enjoy them or be happy with thy own.
we dont follow people, we follow teks and education.
good job violet.
--------------------
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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 Spore Printing! [Re: Violet]
#19157784 - 11/19/13 12:13 AM (10 years, 2 months ago) |
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Wtf jmc? A "ton" of "shit" and "detrimental" information? I notice you first cop-out of discussion entirely without pointing out what that is. You say yourself you're about entirely new to this, only reading things on the board and starting to grow with first pins, yet feel yourself fit to judge? Find a single bit of info that's "shit" or "bad", go on and find it, and we'll point you to the many places where it is an established known. Go on, find all that horrible and wrong stuff that RR and these other peeps must have missed.
However, far be it from me to deny strangers the right to type their 2¢ wherever they want on the internet.
Your statements about me and Anne are ridiculous and behind the curve:
Quote:
FooMan said: FYI- anne and Violet aren't the same person. I've checked and there is no doubt that they are 2 different people in two very different locations. You may not agree with their methods, but please don't falsely accuse them of being puppets of each other.
Aaaaaaanyways.....
Weeks later, I'm still getting 4th-5th flushes from bottom-watered multispore rice pucks, made mainly for the selection of pins for culture isolation
     
I have pretty small 'needs' as far as fruits are concerned, so honestly this culturing alone has had more accidental grow power than I'd expect PFtek to put out on purpose, and with the ease of neglect tek, all the while offering up opportunity for clear visual of ideal genetics.
I'm starting to test some of the isolated clone cultures, and it's looking great...!
-------------------- 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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bowserk
Strangler



Registered: 02/15/13
Posts: 82
Last seen: 3 years, 4 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19158188 - 11/19/13 03:39 AM (10 years, 2 months ago) |
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Thank you for this, this is a lot of great info.
--------------------
Trade List
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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: bowserk]
#19158359 - 11/19/13 05:32 AM (10 years, 2 months ago) |
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Hey now, I started off with a compliment!
Tons of good info here!
Keep up the work!
--------------------
The ego is nothing other than the focus of conscious attention.
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BigGreenMat
Stranger

Registered: 11/04/13
Posts: 161
Last seen: 3 years, 3 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spore Printing! [Re: Violet]
#19159311 - 11/19/13 11:50 AM (10 years, 2 months ago) |
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You are fast becoming my favorite poster! Keeping this thread marked!
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WillSolvem
Odd-Hand



Registered: 04/24/12
Posts: 1,519
Loc: Chapter 26
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spore Printing! [Re: BigGreenMat]
#19159724 - 11/19/13 01:40 PM (10 years, 2 months ago) |
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I like glass and the use of sfd for sterile fae, I spelled it out here 10 months ago here:
http://www.shroomery.org/forums/showflat.php/Number/17459279/page/2
I must say it's a fine write-up
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AMU Q&A Thread because questions deserve answers.
Edited by WillSolvem (11/26/13 05:33 AM)
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nn-IlliniSpiralDMT
Maniac


Registered: 07/19/12
Posts: 380
Last seen: 7 years, 3 months
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Re: Culturing Tech • Agar containers, "Grain Petri" Tek, Invitro growing & Sterile Spores! [Re: Violet]
#19161419 - 11/19/13 07:19 PM (10 years, 2 months ago) |
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Quote:
Violet said: 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!
OWNED
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: Culturing • STERILE SPORES [Re: JMcDoogle]
#19163435 - 11/20/13 03:09 AM (10 years, 2 months ago) |
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Quote:
JMcDoogle said: After reading this thread, there is a huge amount of useful and mind-expanding information, coupled with a ton of shit, and even detrimental information.
And that's just the opening post !
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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]
#19163853 - 11/20/13 07:32 AM (10 years, 2 months ago) |
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Enough of the bullshitting already. You may want it to be true really bad, especially after being told off, but it's simply not. Seriously. All this talk about "bad info", from newbs and nobodys that don't want to actually point any of it out, is entirely contrived. Just wanna throw sticks and run off pretending a jeered insult is true, like ignorant little children.
If there really were any such stuff, do you not think someone could point it out? Do you not thing RR and these other people would have called it? Your pathetic failed attempts with the "malt" and the "x10" were all you could dig up, troll. Give it up. Don't waste your time posting more nonsense bullshit or spin. Don't post again unless you have something real, which you won't; everything in this thread is clearly an established known. The novelty is in the logical arrangement of material and procedure per ends. Set out to prove me wrong, and you're setting out to prove most of mycology wrong. Good luck with that.
-------------------- 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 (11/20/13 07:39 AM)
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