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Invisibleagar
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Spores, G2G, Liquid Culture, Cloning & Colonization Time. * 1
    #3462220 - 12/08/04 12:17 AM (12 years, 5 days ago)

This is the LIFE CYCLE of a mushroom (in nature).


A single mushroom spore contains a half set of chromosomes. When a single spore is in introduced into proper conditions, it germinates. Once germinated, it then sends out a mass of fine thread-like filaments searching for compatible filaments from other germinated spores, doing the same thing. They meet, then (if compatible) mate.

Once mated, the genetically complete cell structure create hyphae. Hyphae filaments is what forms mycelium that is capable of eventually fruiting out as a mushroom. Mycelium has a very high surface area to mass ratio due to the structure of the hyphae, which allows mycelium to absorb large quantities of nutrients from its surroundings after secreting digestive enzymes and digesting its food outside of its body. The ability to intake large quantities of nutrients is a prime reason for rapid mycelia growth.



So, when you inoculate multi-spore solution into a growing medium (grains) in a jar, or bag, they have to germinate, grow, search out & find compatible spore(s), doing the same thing, then mate. Once mated, the resulting mycelium (now genetically capable of fruiting), as rapidly (as conditions permit) colonizes the nutrient medium surrounding it, and under good conditions can eventually form mushrooms, capable of sporulating, thus repeating the life cycle.

SLOWEST: Multi/spore inoculation to a pre-sterilized grain medium.
Does it naturally, and takes the longest, given the extended process.



Grain To Grain (G2G) Transfers


VERY RAPID. Once you have a fully colonized grain jars, or bags, that colonized grain can be manually transferred to pre-sterilized uncolonized grain in fresh jars, or bags. Colonization is very rapid, as you have introduced 100+ fully mated, and colonized contact points, into a fresh grain nutrient medium. G2G transfers is a very rapid method of spawn propagation, and can be accomplished under very aseptic conditions (see trailing G2G method).

THE FASTEST. Liquid culture from spores, colonized grain, or tissue is the FASTEST, (IMHO). Spores germinate & mate rapidly, or tissue / grain is already mated, and the mycelium produced is contained in very mobile solution. (Liquid culture, to succeed requires sterile techniques, as nutrient solutions are very contamination prone). I have left out agar / plate / slurry work as it is not something a novice should immediately attempt.



G2G means Grain to Grain transfers.
Once you have learned to prepare sterilized spawn pint or quart jars of bird seed, rye or grains & fully colonize them with mycelium. You can easily propagate a single colonized quart jar of that into about 20 more via G2G transfers.

Jars propagated via G2G transfers generally colonize under optimal conditions 100% in 10 to 14 days . Given that, spores do not have to germinate, as what is transferred to freshly prepared jars is active mycelium on fully colonized seed or grains.

The method is to prepare fresh jars, just as you would to inoculate via a spore syringe (soak seed, rinse, drain, load, apply filter disk & PC). Excepting, rather than inoculate the fresh jars with a syringe. You transfer grain from a colonized jar to fresh uncolonized jars. The procedure is simple & only requires common sense, minimal preparation, a long stout clean stainless steel spoon & the cleanest personal hygiene and the smallest uncarpeted working place you can muster.

Prepare the smallest cleanest uncarpeted room you have (generally, a bathroom). In the following manner. Clean the room as best you can, getting rid of any dirt, dust, mold or mildew. Remove any cloth hanging anywhere. Spray Lysol on everything, everywhere & wipe it down. If you have any HEPA type air filter unit? Place it in the room & run it for at least 1 hour. Running a HEPA is preferable, but, if you don't have one. You can usually manage without it

Wipe your fully colonized jar of grain & your fresh jars down, with a clean Lysol sprayed rag. Place those in the room, on the counter top, or whatever flat working surface you intend to use. Wear freshly laundered clean cloths. If you have a face mask (preferable) wear it. If you have a shower cap to cover your hair (preferable), wear it. Enter the room, spray Lysol around (again) run the HEPA for a few minutes (if you have one). Then, turn it off. Spray your hands & arms with Lysol & wipe dry.

Unscrew the lid off the colonized jar. Leave the internal filter disk or filter material in place covering the content. Unscrew the lid on a fresh jar, leaving the internal filter material in place. Remove the filter material from the colonized jar & dig up about ? of content, as it will be colonized into a solid mass. Spoon out 2 table spoons full & transfer them to the uncolonized jar, by lifting it's filter up & spooning them in. Replace the filter material on the fresh jar IMMEDIATELY after spooning in the colonized material.

Repeat this same process as many times as you have fresh jars to transfer to. Once done. Screw the fresh jars lids on tight. Cover the outside of the lids of the fresh jars with a double layer of alcohol swabbed coffee filters, or tinfoil & rubber band them down. Shake each fresh jar to spread the colonized material throughout it. Place your fresh jars in a dry, dark, warm place (preferably between 78 & 85 F), and allow them to colonize in peace & quiet. G2G transfer & shaking jars batters the mycelium. It takes it a day or 3 recuperate from that shock. There is no need to shake G2G jars more than once. As, doing so will only slow colonization, rather than speed it up.

The various lid filters you see were experimental.

The very BEST ADVICE I can give anyone who wants to grow mushrooms(beyond cakes)IS: Buy an All American PC, the bigger the better. Build or buy a hepa filter laminar FLOWHOOD.Buy & use BAGS. Buy a Ph test probe. Learn COMPOSTING :thumbup:


Edited by agar (12/08/04 01:07 AM)


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InvisiblePrisoner#1M
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3462302 - 12/08/04 12:36 AM (12 years, 5 days ago)

that was a good refresher for us old timers... excellent info for the new guys.

soemthing I've always admired about you is how thourough your posts are and the
innovative designs you develop for convienience and functionalty....


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Invisiblesmellycat
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: Prisoner#1]
    #3462735 - 12/08/04 02:19 AM (12 years, 5 days ago)

Ooooo maybe i'll actually try G2G now that I know how... :smile: thanks.


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Offlinemushrea
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: smellycat]
    #3463547 - 12/08/04 09:49 AM (12 years, 5 days ago)

Great info


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OfflineMushroomFriend
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: Prisoner#1]
    #3463557 - 12/08/04 09:55 AM (12 years, 5 days ago)

Yeah this is one of the experts who has not much to take from the shroomery, but he brings and gives A LOT! :thumbup:

Agar you rock.


--------------------


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OfflineSilven
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3463559 - 12/08/04 09:56 AM (12 years, 5 days ago)

Amazing agar. :thumbup:

Worth reading more than once for some of us.


--------------------
Born: 10/31/83, which makes me a Scorpio.

1) Scorpios are the most highly sexed of all the signs of the zodiac.
2) Scorpios are prone to excesses: booze, drugs, sex, bad puns, etc.
They usually exploit the weaknesses of others, who fall victim to
their capacity for total lust & sexual abberation.
3) Scorpios possess great intellectual curiosity & creative talent. They
think they are rebels & are arrogant, proud, conceited, and worth every
penny of it.

What do you bring to the table?


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OfflineKaptKid
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: Silven]
    #3463611 - 12/08/04 10:20 AM (12 years, 5 days ago)

Far Out,Man. Really liked the pic of Life Cycle.COOL


Shroom on


--------------------
Child of the 60's, Tripping ever since.


:sun:


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Invisibletripndicular
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3463674 - 12/08/04 10:42 AM (12 years, 5 days ago)

Awesome and very informative as always . :thumbup: :cool: Rating on scale of 1-10 ..." 9.9"
But ...
Do you honestly believe that agar and tissue work should only be preformed by experienced hobbyists ?
Not to be an ass or contradictory to someone I consider an "excellent" choice of mentors ... "I disagree" . I mean agar is more or less the origanl perfected modern day way of getting good healthy cultures for growing "any" species of mush .Granted it is critical that you follow the procedures to a letter and most folks do not have the patience to master the techniques required to keeping "clean" cultures . I only preach this because when I first got into the hobby that was my only choice (I knew of none of the other teks till I found the shroomery  :thumbup:)I jumped in head first , have had great success (maybe just lucky  :wink:)with "minimal"(3%or less) contam problems,and have none of the fancy tools many claim are a neccessity (i.e. flow hood ,glove box ).It just concerns me that this for the "exp only" phrase will spook many away from trying and IMHO be successful at it , maybe even better success than the newer , faster , "popular" teks .
Just a point to keep folks on theirs toes , and not affraid to try it . :wink:

Now for a question about generations of culture .
"WE"(hobbyists in general) do not carry gens of g2g more than 3 gens due to the genetics start to get weakened and degraded .
Do you consider the first petri dish with an "issolated" strain as gen 1 , then the first inoced jar of grain as gen 2 , and then the next as gen 3 our last ?
Or do we start with the very first g2g jar as gen 1 , allowing us 2 more transfers before reaching that final gen 3 ?
Or do you call the first jar inoced with agar wedge as a "master" , then gen 1...2...3...
Sorry if my question sounds as if it going in circles here , I hope it makes sense (kinda prooves I am far from being as wise as you  :wink:)
Again forgive me if it sounds as if I doubt you , I  do not  doubt you and your exp in any way , just wanted to make point that agar is IMO not as "exp only" friendly as most come off sounding . And thank you in advance for answering my gens question , adding to my exp hopefully .


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


Edited by tripndicular (12/08/04 10:46 AM)


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Invisibleagar
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: tripndicular]
    #3465964 - 12/08/04 06:56 PM (12 years, 5 days ago)

I don?t recommend agar plate wedge & slurry work to novices (right off the bat), simply because to do it very effectively (emphasis added) requires (learned) sterile techniques, a very good GLOVE BOX, which are a PITA to work in. Or, a hepa filtered laminar FLOWHOOD (which are optimal & comfortable to work in), but cost much more to build, or buy.

To incubate and/or store agar cultures in optimal conditions requires an effective incubator, with accurate temperature control. Optimal bulk slurry (or liquid culture slurry) work requires a magnetic mixer plate, stir bars & glassware and/or an Eberbach type laboratory blender, which are expensive.

Not that all that can?t be done with various ghetto TEK?s, it can. But IMHO, they often result in a fair amount of failures, which are disconcerting to a novice.

Learning to crawl, before one walks is wise. That way one doesn?t stumble, or fall - as often. It?s better (IMHO) to start PF style, learn aseptic techniques, needle work, get a successful PF grow (or 3) under your belt.

Then learn PRINTING. Once you are at that stage (having actually completed a grow, or 3), & have your own spore supply, you can decide to go as far beyond stage - as you wish & how much you are willing to invest in the equipment to do that EFFECTIVLY.

Everyone?s personal circumstance DIFFERS. The information in the massive number of TEK?s does to. There is so much Bunko information floating in cyber/space, it is confusing to a novice. One must ferret out the good / factual information, learn what works for them, then expand on it - if so inclined.

I started with BULK, because I had the inclination, patience, time, room, privacy & means. Without all that, novices face a lot of stumbling blocks. Proof of that is seen every day, in numerous "what now" posts on this very board.

As for generations, IMHO, MASTER GRAIN JAR, is from spores. Generation (G) 1, is G2G from Master Jar. G2 is G2G from G1, G3 is G2G from G2, so on & so forth.

IMHO, Agar MASTER PLATE is from spores (Or, tissue = clone). Agar wedge or slurry transfer to grain spawn is G1. G2G from that is G2, so on & so forth.

Tissue (clone) to agar plate is Master Plate. Agar wedge slurry out of Master Plate into grain jar is G1. A week later, agar wedge slurry from same Master Plate is still G1.

CULTURE STORAGE:
Instead of trying to keep cultures alive, there is a better way: In sterile distilled water, with no food, oxygen, or minerals. This method was in use almost 60 years ago, but was apparently lost due to lack of communication.

Sterile water was first used to preserve cultures by Castellani in 1939 (2). Since then, many scientists have used this method; McGinnis et al. in 1974 (3) and Odds in 1991 (4) reported that they were able to maintain viable cultures for more than three years, without degradation.

This technique satisfies many different interests: Castellani's pathogenic fungi, Odds interest was in pathogenic yeast, and McGinnis' interest was in a wide range of fungi, yeast, and bacteria. The distilled water preserved all of them.

Storage Method:
Obtain small vials from your laboratory supplier and fill them about half full (about 3mL) with distilled water, loosely cap the vials and sterilize them in a pressure cooker for 30 minutes @250 F. Half vials or test tubes with screw caps would also work well.

About six milliliters of sterile distilled water is pipetted aseptically into a freshly growing culture. The fragments of hyphae are dislodged by lightly scraping the aerial growth with the same pipette, and the resulting suspension is withdrawn and transferred to a sterile glass vial.

Put plenty of inoculum into each vial to insure success. Screw the lid on tight and wrap Parafilm around the top of the vial to make sure it is airtight. When you come back in a few years, you would not want to find that the water had evaporated.

Store the vials at room temperature away from direct sunlight. A wall cabinet is an excellent place. If conditions deteriorate, and the room should become extremely hot, the vials can be refrigerated, but that is not normally necessary.

In the distilled water environment, the mushroom culture enters a dormant state, and it is held in stasis.

The Rude Awakening:
Under aseptic conditions, simply dip a sterile loop into the vial, and streak the mycelia-rich water onto an agar plate. It will start to reanimate on being in a nutrient source and oxygen.

References
(1) M.D.Graham, "A Simple,Practical Method for Long Term Storage of Yeast", Brewing Techniques 5, March/April (1997), pp 58-62
(2) S.Castellani,"Viability of Some Pathogenic Fungi in Distilled Water", Journal of Tropical Medicine and Hygiene 42, pp 225-226 (1939)
(3) M.R.McGinnis,A.A.Padhye,and L. Ajello,"Storage of Stock Cultures of Filamentous Fungi,Yeasts, and Some Aerobic Actinomycetes in Sterile Distilled Water", Applied Microbiology 28, pp 218-222 (1974)
(4) F.C.Odds, "Long Term Laboratory Preservation of Pathogenic Yeast in Water",Journal of Medical and Veterinary Mycology 29, pp 413-415(1991)

~~~~~~~~~~~~~~~~~~~

Vinegar and Hydrogen Peroxide as Disinfectants
by Judy Stouffer, B.S., M.S., SFO

You can make your kitchen a cleaner, safer place and fight bacteria, without exposing yourself and your family to toxic chemicals that also damage the environment. You can use a simple safe disinfecting spray that is more effective than any of the commercial cleaners in killing bacteria. As a bonus, it is inexpensive!

Susan Sumner, a food scientist at Virginia Polytechnic Institute and State University, worked out the recipe for just such a sanitizing combo. All you need is three percent hydrogen peroxide, the same strength available at the drug store for gargling or disinfecting wounds, and plain white or apple cider vinegar, and a pair of brand new clean sprayers, like the kind you use to dampen laundry before ironing.

If you're cleaning vegetables or fruit, just spritz them well first with both the vinegar and the hydrogen peroxide, and then rinse them off under running water.

It doesn't matter which you use first - you can spray with the vinegar then the hydrogen peroxide, or with the hydrogen peroxide followed by the vinegar. You won't get any lingering taste of vinegar or hydrogen peroxide, and neither is toxic to you if a small amount remains on the produce.

As a bonus: The paired sprays work exceptionally well in sanitizing counters and other food preparation surfaces -- including wood cutting boards.

In tests run at Virginia Polytechnic Institute and State University, pairing the two mists killed virtually all Salmonella, Shigella, or E. coli bacteria on heavily contaminated food and surfaces when used in this fashion, making this spray combination more effective at killing these potentially lethal bacteria than chlorine bleach or any commercially available kitchen cleaner.

The best results came from using one mist right after the other - it is 10 times more effective than using either spray by itself and more effective than mixing the vinegar and hydrogen peroxide in one sprayer.

Science News August 8, 1998; Vol. 154, Issue. 6; pg. 83-85

EFFECTIVE INCUBATOR & GROW CHAMBER

A broken freezer can be acquired FREE from most appliance repair places.

EFFECTIVE HEATING METHOD
WATERBED HEATER & CONTOL


ANOTHER EFFECTIVE HEATING METHOD

Contol & wiring from electric blanket

FRUITS OF LABOR


--------------------


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Invisibletripndicular
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3469415 - 12/09/04 10:06 AM (12 years, 4 days ago)

Book ... book ... book ... I want a first edition , signed , sealed and delivered too .... :wink:

Thanx for the detail master agar . I am so full of info I am about to explode  :eek:

Your ideal gens naming may be why I have run into a few situations where a species (one in particular-Malabar)would not produce spores , especially in the second and third flushes . From what was being named gen 3 (actually 4 by your description). Gonna play with that some more just to learn .
IMO from genetics getting a litte degraded , I also considered the possibilty of mutatons , due to Mals being genetically selected for warmer climates by the origanal mycologist\vendor or just simply a quirk in them .

I never knew we could long termstore in just sterile water , I like that alot , may even try it out .

I know all about vinegar to clean any and allareas of home , in fact I try to promote it , but many doubt the power of vinegar  :frown: :confused: In fact today is scrub the ole tile floor with the 2:1 vinegar water solution . :wink:


Thanx again as always I am enlightened to learn from you ! :cool: :thumbup:


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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Offlineqwon
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3469951 - 12/09/04 12:38 PM (12 years, 4 days ago)

Liquid Mycelium = the way to go. Beats gTg anyday. Fastest afoaf had a completely col. jar = 7 days. Back to back, did gTg with 1 1/2 tbsp. broken myc into 1/2 pint = 1 week not yet 3/4 col. Same day did Liquid Myc. 4 days = slightly the whole jars are becoming whiter by the day...
thx agar


--------------------
'I'm the commander see, I don't need to explain, I do not need to
explain why I say things. That's the interesting thing about being the president.
Maybe somebody needs to explain to me why they say something,
but I don't feel like I owe anybody an explanation.'
- George W. Bush


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Offlinescatmanrav
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: qwon] * 2
    #3470686 - 12/09/04 03:22 PM (12 years, 4 days ago)

I must also disagree about only novices not working with agar. I started out months ago in this hobby with everyone going "PF TEK! PF TEK!" when I asked what route to go. I didnt listen and instead ordered agar and bought rye and started there. I had tremendous success with both inculding isolating a syringe I got that was contamed..for fun. I tried it in 6 quarts and 5 petris, all grew green, 3 other strains all using 6 jars each and 5 petris each and not a single contam other then that. After a few attempts and transfers I learned how to do it. First I tried taking a bit of germinated myc from grain jars (couldnt get the myc to germinate worth while on the agar, green took it to fast) into other grain jars but it just perpetuated the mold..moving to agar though I got some nice clean growth and after a few transfers I have a clean (and agressive) GT specimen from a dirty one. This was in my first 3 weeks or so of being in this hobby. I learned alot from it and I wouldn't tell others to hold back if they are comfortable.

I do however have extreme sterile conditions (no glove box or flow hood but I strip my bathroom to barebones solid floor, no cloth and run 2 HEPAs in there for a few hours prior to work and clean it well) that I use all the time for g2g transfers and agar work with very very little contam rate (like 1 or 2 out of every 50-100 g2g transfers or agar plates).

Also you said that liquid culture is the fastest? IME grain to grain transfers have been faster then liquid cultures...

Very good info though.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Invisibletripndicular
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: scatmanrav]
    #3470721 - 12/09/04 03:29 PM (12 years, 4 days ago)

Remeber shutting down ventalation system to house helps , I hope I'm not assuming you did not , but most folks forget to prevent much air flow from getting into work place helps alot . I think kitchen is best for me , I do not crap in there  :wink:

Thanx for the back up ! Like your avatar says ......

"THINK OUTSIDE THE BOX !" I love that !

Once you go agar , you'll never go back IMHO !

PS are we bakers or mushroom makers I always say !


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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Offlinescatmanrav
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: tripndicular]
    #3470769 - 12/09/04 03:40 PM (12 years, 4 days ago)

I dont shut off the air flow to my house because the bathrooms doesnt have a central vent..I dont open the door to move stuff in or out (like myself when Im ready to go in after hours) with the air kicked on and I spray lysol in front of the closed door and open it quickly and walk through it.

I also do this after a shower and dont put on anything other then clean boxers, gloves, and a paint mask.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Offlinespliffmasta
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: scatmanrav]
    #3470830 - 12/09/04 03:53 PM (12 years, 4 days ago)

There is no way that liquid culture is faster than a grain to grain transfer.

1) The myc in a liquid culture is very weak and takes a little bit to recuperate.
2) With G2G you're allowing for more spawn points with stronger, more ready-to-grow myc.

A G2G transfer is almost like starting at the point when you use a liquid culture and give it its first good shake.


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Invisibletripndicular
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: scatmanrav]
    #3470861 - 12/09/04 03:57 PM (12 years, 4 days ago)

Sexy , boxers , mask ,gloves ........ :wink: :tongue:
Get a hair cap to , lots of shit falls off head no matter how clean you are !


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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Offlinescatmanrav
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: spliffmasta]
    #3470998 - 12/09/04 04:15 PM (12 years, 4 days ago)

Quote:

spliffmasta said:
There is no way that liquid culture is faster than a grain to grain transfer.

1) The myc in a liquid culture is very weak and takes a little bit to recuperate.
2) With G2G you're allowing for more spawn points with stronger, more ready-to-grow myc.

A G2G transfer is almost like starting at the point when you use a liquid culture and give it its first good shake.




Right and I know agar has said before in a post that G2G is the faster way in his opinion..Im pretty sure he said that..thats why I think that was just mis typed..didnt notice it being mentioned when scanning through responses though so thought I would mention it.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Invisibleagar
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: scatmanrav]
    #3472588 - 12/09/04 09:31 PM (12 years, 4 days ago)

Quote:

scatmanrav said:
Quote:

spliffmasta said:
There is no way that liquid culture is faster than a grain to grain transfer.

1) The myc in a liquid culture is very weak and takes a little bit to recuperate.
2) With G2G you're allowing for more spawn points with stronger, more ready-to-grow myc.

A G2G transfer is almost like starting at the point when you use a liquid culture and give it its first good shake.




Right and I know agar has said before in a post that G2G is the faster way in his opinion..Im pretty sure he said that..thats why I think that was just mis typed..didnt notice it being mentioned when scanning through responses though so thought I would mention it.



~~~~~~~~~~~~~~~~~~~
Grin???..
Okay respected members?? you called me out on this?.

So allow me to explain from my point of view. :laugh:

G2G is the fastest STRAIGHT FORWARD METHOD.

One step beyond that & not so straight forward is liquid culture.

OPTIMAL liquid culture is the  FASTEST  & here is why (keyword OPTIMAL).

FIRST OFF, everyone?s "liquid culture" RECIPE differs, simply because they use "whatever they can get", that they think & hope will work. Some use Karo (corn sugar syrup), some use honey water, some use light dry malt, some use powdered dextrose, nutritional yeast, potato water, rye water, peptone, soytone, rice water, near beer, combinations of all of the above and/or any number of whatever else comes to mind - after "bong hit 4".

So, all liquid culture solution nutrient mediums ARE NOT EQUAL.

The container type, size & lid everyone uses to hold &  any liquid culture in - ARE NOT EQUAL.

Most just use whatever glass jar & lid that is handy - period. Others use lab grade glassware (or equivalent glassware), with devices to hepa/ulpa filter aerate, stir & aspirate the liquid culture with.

SIZE MATTERS (:tongue2: & women lie about that - fact)

You obviously get MORE MYCELIUM SOLUTION in a 1 gallon container, than you do in a half pint jar.

So, liquid culture containers DIFFER GREATLY. :wink:

INCUBATION:

Some just use a closet shelf & plastic box for incubation. Other use much more regulated incubators.

INCUBATION procedures DIFFER GREATLY.

Then there is what you use to SEED the liquid culture with.

Some use spores, some use agar isolates, some use colonized grains, some use mushroom tissue, etc.

So, there are innumerable VARIBLES in liquid culture recipes, preparation, containers, incubation, aspiration & dispersal methods .

If you use an agar plate isolate to seed liquid culture with, you know it?s clean, mated & you pick the most aggressive growth. That goes into your nutrient solution, is incubated & colonizes.

Under OPTIMAL CONDITIONS mycelium will be exponentially multiply from a tiny fragment, to a very - very large biomass VERY - VERY RAPIDLY.


QUART JAR EXAMPLE


When this is accomplished, the resulting mycelium is so thick, you cannot even aspirate it through a normal 18 gauge syringe needle, as It will instantly PLUG the needle.

You can use LARGE CAPICTY SYRINGES (60 ml - cc) & large bore plastic needles to work out of a jar.

Or use use a self filling syringe system:



Or,  a stirring / cutting mechanism  inside the jar, or a Eberbach type lab blender.





Once done in volume (2+ quart, 2+ liter, per container) you have a massive amount very FINE SLURRY mycelium biomass to inoculate with.

If all is done in OPTIMAL fashion, whatever you inoculate, will colonize FAST - because you are introducing  MILLIONS  of mycelia contact points, rather than hundreds, as in G2G. 

You can even inoculate straight into an OPTIMAL substrate & it will colonize rapidly (but - that is another story).

So, there is a GREAT DIFFERANCE between someone with a half pint jar, with a few wisps mycelia in it, than a 3/4 gallon batch of OPTIMAL mycelia slurry,  :tongue2: thick as a dairy queen milkshake :tongue2:. :thumbup:


--------------------


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Offlinescatmanrav
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Registered: 05/08/04
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Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: agar]
    #3472915 - 12/09/04 10:13 PM (12 years, 4 days ago)

I see your point but I'm still not so sure.

So if you take your optimal culture and knock up a jar..its made with an isolate and is thick as hell..it colonizes incredibly quick..moreso then most grain to grain transfers..but then you take two spoon fulls of that jar into a new jar and thats the same great isolate but probably more myc (solid 2 spoonfulls is quite a bit of volume). I relize it could be fewer contact points but I'm also inclined to think that the liquid myc clumps to the grains rather well (none of my solutions have been thick enough for seeing so this is guessing) and wouldnt really spread all over so much after being squirted in. What would you say would happen and how many cc's would you use to knock up the jar. Then how long would jar 1 take to colonize and how long for jar 2 with the g2g to colonize? Just a guess so I could tell a difference.

I do use a light karo solution (though more then everyone else says). Everyone says a teaspoon of karo per 100ml of water but I like a tablespoon or two. Malt is more difficult to get ahold of for me..Karo was 3 for 99 cents on sale and I got enough for a while.... Plus like I said (I think I did in this post) I'm not so concerned with colonization times since fruiting takes much longer then colonizing.

Nice stuff I'd like to get into someday..not that much excess funds to play with as of yet though.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


Post Extras: Print Post  Remind Me! Notify Moderator
Invisibletripndicular
My Minds Eye IsRhizomorphic

Registered: 08/25/02
Posts: 2,791
Loc: Bowels of HELL
Re: Spores, G2G, Liquid Culture, Cloning & Colonization Time. [Re: scatmanrav]
    #3472996 - 12/09/04 10:25 PM (12 years, 4 days ago)

agar is in another league guys , never doubt his methodologies , he will just blow your mind with more knowledge that you can only hope to comprhend !
How long ?????? To many "x" factors involved !

Awesome detail on what "true" liquid "slurry" is old man .
My head sure is swimming in it right now ! :wink:
I love that auto injector .......sweeeeeeeeeeeeetttttttttttt !


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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