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My first project: Sub Strain Isolation. Looking for critique on my tek
    #6346826 -

Well, in 2-3 months I'll be moving into a larger apartment with significantly more closet space!  :thumbup: At that time, I plan on setting up a Martha or two, as well as throwing together a flow hood, and dabbling with a few different species of edibles and actives.

In the mean time, I only have one closet to work with, and a simple glove box. My plan is to isolate a potent, fast colonizing, high yielding sub-strain of cubensis. I'll be starting with Koh Samui Super, P.E.S.A, and Cambodian strains.

Each syringe will be used to inoculate 10 pre-sterilized slides of agar in a sterile glove box. While they are incubating, growth will be carefully monitored and recorded. From each strain, the two fastest growing slides that show rhizomorphic mycelium development will be chosen as spawn.

The 6 cultures will inoculate 3 rye grain jars (rye:water at 1.25:.9 ratio) each. The water will be laced with H2O2 to help insure sterility. Inoculation will take place inside a sterilized glove box. While the 18 jars incubate, speed of colonization will be monitored and recorded. They will be shaken as needed to ensure even distribution of spawn.

When the jars are fully colonized, they will be mixed with pre-sterilized mushroom compost at a 1 part spawn: 3 parts substrate ratio.  This will take place inside a sterile glove box, and the substrate will be placed into plastic trays covered with duct tape to keep out light. Speed of substrate colonization during incubation will be monitored and recorded.

After the trays are fully colonized, they will be cased with ½-5/8” of 50/50 casing mix, and misted thoroughly with H2O2/H2O, and left covered to incubate for 3 days. After 3 days, they will be checked. If myc is poking through, they will be patched as needed, and misted, then put into the FC.

Time from casing to pinning will be recorded. Fanning will take place 4-6 times a day, with a light misting after each fanning until pinning. After pinning, fanning will be cut back to 2-3 times a day, with light misting only if humidity drops below 90%.

Harvest will take place just before the veil tears. The largest fruit body from each of the 6 sub-strains will be isolated for cloning. The tissue sample will be dropped into a water/H2O2 mixture for sterilization and storage while the harvest is processed. The harvest will be divided based on sub-strain, and each of the 6 sub-strains will be ranked in terms of colonization speed, time from casing to pinning, and overall yield.

After the harvest is dried, I will recruit some willing and experienced volunteers to participate in a blind study to subjectively rank the potency of each of the 6 strains.

Based on this subjective potency data, combined with the previously collected data on growth speed, I will work to create the “ideal” sub-strain. The earlier tissue samples will be used to create clone mycelium of the largest fruit bodies from the 6 original sub-strains of peroxide laced agar.

After the potency rankings have been completed, I will cross the 2 most potent sub-strains with the two fastest growing. I will then repeat the procedure outlined above for these new sub-strains, to attempt to further enhance my stock.

My goal is to end up with the best sub-strain possible 2-3 months down the road so I end with the best quality mushrooms possible from my Martha setup. I deliberately selected strains that are supposedly both potent and quick/rhizomorphic colonizers.

What do you all think of the plan I’ve outlined here? I’m a complete noobie to mushroom cultivation. All of this is based off of the tremendous wealth of information available on this site and these boards. I’ve been lurking and researching for a good 2 months while I’ve put together my plans, and the above is what I’ve come up with. Any criticism would be greatly appreciated


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Knock the world right off its feet and straight onto its head.

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: Cubeculture]
    #6346947 -

Quote:
The 6 cultures will inoculate 3 rye grain jars (rye:water at 1.25:.9 ratio) each. The water will be laced with H2O2 to help insure sterility.


I don't know if the H2O2 is necessary in the water for this part, but wait for an old hand to chime in.

Second, why are you going to dry the harvest before consumption? Munch that shit fresh and trip balls! (I hope that you will be one of the experiencers as well, gotta eat the fruits of your labor!)

2-3 months seems kinda rushed. I don't know that you'll be able to complete it in that time, but it depends on how many generations you're going to do. You'll be able to get thru one or two grows, but whether that's enough for the isolation you intend to do, I don't know.

Quote:
Based on this subjective potency data, combined with the previously collected data on growth speed, I will work to create the “ideal” sub-strain.


Are you aware of how difficult it is to successfully hybridize two strains of mushroom? It is nothing like cannabis, which is very easy to cross. It is possible, but very difficult. I would suggest contacting RogerRabbit on this subject.

Also, you say that you want to harvest before the veil tears (which I do, and agree with), but this will not result in maximum size. Granted, they probably won't get much bigger after that point, but play around with it.

My last point: WHY AREN'T YOU USING CREEPERS?!?!? I have grown these. They colonize very quickly with rizomorphic mycelium. The are very contaminant resistant. They are prolific fruiters with med-large fruits. Lastly, THEY ARE VERY POTENT! Last week, I was able to harvest some, and to test their potency, ate 6g wet. The result? I can't even explain it. Needless to say, VERY POTENT. I would recommend considering this strain in addition to/replacement of one of the others.

Finally, GOOD LUCK! It seems you have done much research, and intend to be very cautious. I hope your grow goes well, and hope that you will enjoy your fruits!

Edited by MistaUNGA (12/08/06 03:30 AM)

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: MistaUNGA]
    #6347152 -

If I recall correctly, H2O2 breaks down under the heat of PCing so there is really no need to use it. There's also really no need to use it on casing unless you're trying to combat cobweb. But, fresh air will keep that at bay.

I'd also suggest not spreading spores to 10 dishes. You'll have plenty of genetic variation on 1. Just pick a couple sectors on that one, and isolate each one.

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: BataviaVakereli]
    #6347199 -

you ate 6 grams wet and you say its very potent? isnt that like eating .6g dry? that doesnt seem like enough to get a body buzz from.


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They who can give up essential liberty to obtain a little temporary safety deserve neither liberty nor safety.
Franklin's Contributions to the Conference on February 17 (III) Fri, Feb 17, 1775

Believe nothing, no matter where you read it, or who said it, no matter if I have said it, unless it agrees with your own reason and your own common sense. - Buddha

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: BataviaVakereli]
    #6347214 -

BataviaVakereli said:
There's also really no need to use it on casing unless you're trying to combat cobweb.  But, fresh air will keep that at bay.




:thumbup: :thumbup:

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: Blutjager]
    #6347684 -

You'll need to learn to crawl before you walk my friend. What you've described is not strain isolation at all. Concentrate at first on getting a harvest and learn the basics such as grain preparation. Forget the peroxide, it's for cobweb mold, not grains.

Each drop of spore solution from a syringe will have up to a few dozen strains worth of spores. Strain isolation is allowing the plate to grow out for a few days, then making ten or more transfers from the leading edge of the growth to new plates. The second set of plates should begin to show some sectoring. Transfer each sector to new dishes, and continue to repeat the process until you have non-sectoring mycelium in each plate. At this time, transfer part of each dish to well marked quarts of rye to grow out, and put the rest of each plate in the refrigerator until the rye has grown out and fruited, then tested for the qualities you desire. When you find the strains you like, you can then go back to the refrigerator, and pull those plates out to make master slants with, that can then be stored and used for many years.

Don't fall into the trap of thinking the 'prettiest' or most rhizomorphic mycelium is the best. Fruit out each substrain you isolate to determine quality.

Two or three drops of solution from a single syringe can easily use up 100 or more petri dishes by the time you isolate each substrain out. for this reason, I purchase three and four compartment petri dishes, so each dish can have multiple cultures, saving a lot of money.
RR


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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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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: RogerRabbit]
    #6347728 -

Thanks for all the feedback folks.

Perhaps I would be better served by trying to perfect my pinning strategy before I work on my strain's genetics...


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Knock the world right off its feet and straight onto its head.

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: DrB]
    #6348014 -

DrB said:
you ate 6 grams wet and you say its very potent? isnt that like eating .6g dry? that doesnt seem like enough to get a body buzz from.


Hey don't ask me why. It was only supposed to be a little test of potency, I was not expecting to trip at all. I expected a SLIGHT body buzz. I got a complete mind fuck.

Anyways, just do like RR said. He won't steer you wrong!


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:gc:
Madtowntripper said:Or just give her a cloroform soaked rag and tell her it's ether!

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: Cubeculture]
    #6348037 -

1. Buy largest PC you can afford.
(All American Brand is optimal)

2. Buy spore syringes from a Shroomery vendor.

3. Buy a couple cases of quart Mason jars.

4. Learn wbs/grain spawn making in jars.
(includes soaking, jar lid gas exchange provisions & injecting)

5. Figure out wbs/grain jar incubation method.

6. While spawn jars are colonizing. Build fruiting chamber.
(learn about impeller coolmists & FAE provisions)

7. While spawn jars are incubating. Find free horse manure source.

8. Learn h/poo substrate preparation method.

9. Spawn h/poo substrate trays.

10. While trays are incubating, Learn Casing preparation methods.

11. Once trays are colonized, add casing cover & incubate more.

12. Fruit trays.

13. Learn printing methods.

14. Buy a Nesco brand dehydrator.

15. Enjoy the fruits of your labor.

16. Improve everything & go again, only better.
(glove box & flow hood)

17. Spread the  :mushroom2: :mushroom2:MAGIC :mushroom2: :mushroom2:.


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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: agar]
    #6348246 -

Spores are in the mail! Only thing I'm having trouble tracking down locally are jars... I've gone to 7 different supermakets, and none of them have any in stock. It looks as though I might have to order them too, but if that's the biggest road bump I hit, I figure I'm not doing too badly.

An All American PC is next on my list after the jars  :thumbup:

I'm not sure I'll be able to find a free h-poo source, I live in the burbs, and it's quite a drive to the closest pasture. I was planning on going with High Mountain compost.

I've been doing a lot of reading, think I have the basic concepts down, and I think I'll take the advice of all the old hands and start out small before I try anything too ambitious. Thanks for all the advice!


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Knock the world right off its feet and straight onto its head.

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: Cubeculture]
    #6348342 -

LOL,

Way back when.
I started BIG.
(big = grain spawn to bulk substrate)

First thing I learned was about BIG MISTAKES. :rolleyes:

Wise to get SMALL down pat, FIRST.
That makes BIG, a lot easier, later.



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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: agar]
    #6348386 -

Maybe I'll just start with simple cased rye. KISS


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Knock the world right off its feet and straight onto its head.

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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: Cubeculture]
    #6348408 -

Cubeculture said:
Maybe I'll just start with simple cased rye. KISS


Just as simple is 50/50 rye/coir, as bulk substate.
Yeild will be higher than rye alone


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Re: My first project: Sub Strain Isolation. Looking for critique on my tek [Re: agar]
    #6348502 -

agar said:
Cubeculture said:
Maybe I'll just start with simple cased rye. KISS


Just as simple is 50/50 rye/coir, as bulk substate.
Yeild will be higher than rye alone


Something else to add to my shopping list, thanks for the ideas!  :thumbup:


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Knock the world right off its feet and straight onto its head.

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Shop: North Spore Bulk Substrate   Sporeworks.EU Spores for European Microscopy   Myyco.com Isolated Cubensis Liquid Culture For Sale   MagicBag.co All-In-One Bags That Don't Suck   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Original Sensible Seeds Autoflowering Cannabis Seeds   Mushroom-Hut Liquid Cultures

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