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KauaiOrca
Waterman


Registered: 08/12/08
Posts: 3,131
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Re: My invitro techinque [Re: Pastywhyte]
#21953610 - 07/16/15 08:17 PM (8 years, 6 months ago) |
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Quote:
Pastywhyte said: Agreed. Invitro has its merits, I have done a few that way myself 
I think, Invitro, there is absolutely nothing that comes close to how easy Sclerotia is … ultimate set and forget method. I'm trying to perfect a combination of Violet's culturing method with RGS and Muda's grow bottles with a SGFC or if I get it going right, maybe a mini mono for 4-6 bottles.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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jesuisravi
The Old Noob



Registered: 06/24/15
Posts: 260
Loc: Midwest USA
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Quote:
spacechildo said:
Quote:
KauaiOrca said: but they involve a fruiting chamber.
Cron's Growing without a Fruiting chamber
teaches you very well how to read the mushrooms. read the whole thread for lots of good input on how to pull it off.
Thanks for posting this. A breakthrough! I would never had found this using the search engine here--should I blame the engine or the would be engineer? Anyway, an inspiring link. Double to you.
-------------------- Most of my beliefs I acquired from my father and from John Wayne, and anything that wasn't ultra tough and ultra cool was to me ultra embarrassing. In fact, I lived in a state of near continuous embarrassment, never measuring up to the ridiculous standards I had accepted without question, applied to a framework of expectations neither I nor anyone else could meet.--J C Amberchele almost nothing important that ever happens to you happens because you engineer it. Destiny has no beeper; destiny always leans trenchcoated out of an alley with some sort of 'psst' that you usually can't even hear because you're in such a rush to or from something important you've tried to engineer. ” ― David Foster Wallace, Infinite Jest
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Buck513

Registered: 04/17/14
Posts: 5,682
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Re: My invitro techinque [Re: durxo]
#21954651 - 07/17/15 02:46 AM (8 years, 6 months ago) |
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Quote:
durxo said:
Quote:
blackout said: I would skip the alcohol, it can harbour contams. Just cover the lid with foil, and only remove the foil just before you are inoculating, so the injection ports will be sterile.
It will also stop any water getting on your lid and into the containers through the filters. It looks like your lid might have a ridge running around it which will possibly make a pool in the centre if water drips off the top of the PC.
I would also recommend people to drill plastic on top of some wood, the plastic in the pics looks thick and not too brittle, others will crack. I usually drill a small hole and hold a lighter up to it, this smoothens out the edges and enlarges the hole and makes it stronger and smoother.
Yes you could too but the point of the isopropyl alcohol is to kill anything that might be in the needle and its really effective I dont care anymore where I do my prodecures, I do it in open air with the AC on, even in the kitchen where there is mold or with an open windows, and the alhocol doesnt make it trough the injection port, maybe some that might get caught inside the needle but never gave me any problems.
I keep my isopropyl alcohol inside a tincture bottle with a dropper.
So...are you one of those people that believe alcohol kills mold/bacteria efficiently?
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: Buck513]
#21957222 - 07/17/15 05:53 PM (8 years, 6 months ago) |
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Quote:
Buck513 said:
Quote:
durxo said:
Quote:
blackout said: I would skip the alcohol, it can harbour contams. Just cover the lid with foil, and only remove the foil just before you are inoculating, so the injection ports will be sterile.
It will also stop any water getting on your lid and into the containers through the filters. It looks like your lid might have a ridge running around it which will possibly make a pool in the centre if water drips off the top of the PC.
I would also recommend people to drill plastic on top of some wood, the plastic in the pics looks thick and not too brittle, others will crack. I usually drill a small hole and hold a lighter up to it, this smoothens out the edges and enlarges the hole and makes it stronger and smoother.
Yes you could too but the point of the isopropyl alcohol is to kill anything that might be in the needle and its really effective I dont care anymore where I do my prodecures, I do it in open air with the AC on, even in the kitchen where there is mold or with an open windows, and the alhocol doesnt make it trough the injection port, maybe some that might get caught inside the needle but never gave me any problems.
I keep my isopropyl alcohol inside a tincture bottle with a dropper.
So...are you one of those people that believe alcohol kills mold/bacteria efficiently?
Well at least for me it works really well, Ive never flame sterilized anything, I dont know if its my alcohol cause I have a laboratory grade 99% pure, so maybe is that but I dont believe it, Ive seen it many times, maybe Im wrong but I will keep doing it as long as it gives me good results.
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: KauaiOrca]
#21957240 - 07/17/15 05:56 PM (8 years, 6 months ago) |
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I know, Im trying to cultivate truffles but I inoculated my bag with P. Mexicana like 6 weeks ago and nothing, I have the bag in a dark place at about 77-80 fahrenheit but I dont see any truffles, heres a picture.
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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spacechildo
proletarians rise up



Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
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Re: My invitro techinque [Re: durxo]
#21957246 - 07/17/15 05:58 PM (8 years, 6 months ago) |
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how do you know flame sterilizing wont give you even better results if you havent tried it?
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Buck513

Registered: 04/17/14
Posts: 5,682
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Re: My invitro techinque [Re: durxo]
#21957249 - 07/17/15 05:58 PM (8 years, 6 months ago) |
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I mean....its a fact that alcohol doesn't kill all that it needs to. Alcohol sanatizes, heat sterilizes. There is a tremendous difference.
Someone posted this awhile ago in another thread, the facts are right here why you shouldn't use alcohol like that
http://www.cdc.gov/hicpac/Disinfection_Sterilization/6_0disinfection.html
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Buck513

Registered: 04/17/14
Posts: 5,682
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Re: My invitro techinque [Re: durxo]
#21957283 - 07/17/15 06:04 PM (8 years, 6 months ago) |
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Just because you get mushrooms to fruit, doesn't mean it was a success, or that it wasn't contaminated. Many don't seem to realize that
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: My invitro techinque [Re: durxo]
#21957304 - 07/17/15 06:09 PM (8 years, 6 months ago) |
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Quote:
durxo said: I know, Im trying to cultivate truffles but I inoculated my bag with P. Mexicana like 6 weeks ago and nothing, I have the bag in a dark place at about 77-80 fahrenheit but I dont see any truffles, heres a picture.

Bags always look sketchy but I think that looks bacterial. . .
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spacechildo
proletarians rise up



Registered: 01/24/13
Posts: 19,243
Loc: Babylon
Last seen: 6 years, 4 months
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Re: My invitro techinque [Re: Pastywhyte]
#21957318 - 07/17/15 06:12 PM (8 years, 6 months ago) |
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that's majorly bacterial.
but I have no exp with bacteria and stone formation so IDK if that's why
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KauaiOrca
Waterman


Registered: 08/12/08
Posts: 3,131
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Re: My invitro techinque [Re: Pastywhyte]
#21957528 - 07/17/15 07:12 PM (8 years, 6 months ago) |
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Quote:
Pastywhyte said:
Quote:
durxo said: I know, Im trying to cultivate truffles but I inoculated my bag with P. Mexicana like 6 weeks ago and nothing, I have the bag in a dark place at about 77-80 fahrenheit but I dont see any truffles, heres a picture.

Bags always look sketchy but I think that looks bacterial. . .
I agree, that bag doesn't look good. I got a Galindoi bag about that size going exactly 30 days ago and there are carmel and brown colored stones everywhere.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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Buck513

Registered: 04/17/14
Posts: 5,682
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Re: My invitro techinque [Re: KauaiOrca]
#21957546 - 07/17/15 07:17 PM (8 years, 6 months ago) |
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I guess alcohol doesn't make such a great sterilizer after all
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Re: My invitro techinque [Re: KauaiOrca]
#21957600 - 07/17/15 07:32 PM (8 years, 6 months ago) |
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Is possible you maybe didn't inject Mexicana into it, but maybe P.C. Cubensis Mexican?
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: Grey]
#21958734 - 07/17/15 11:38 PM (8 years, 6 months ago) |
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Quote:
Grey said: Is possible you maybe didn't inject Mexicana into it, but maybe P.C. Cubensis Mexican?
Yeah its possible cause I bought the loquid culture on internet.
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: Buck513]
#21958740 - 07/17/15 11:40 PM (8 years, 6 months ago) |
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So you guys think it could be bacterial? I said alcohol does a good job cause when I cultivate shrooms everything looks fine, mycellium looks white and no strange odors.
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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Buck513

Registered: 04/17/14
Posts: 5,682
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Re: My invitro techinque [Re: durxo]
#21958747 - 07/17/15 11:43 PM (8 years, 6 months ago) |
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I've managed to get more than 4-5oz from 1 flush on a contaminated sub.
Looks can be deceiving.
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Re: My invitro techinque [Re: durxo]
#21958756 - 07/17/15 11:45 PM (8 years, 6 months ago) |
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To me, it looks like overcolonized grains by cube mycelium. If it's a 100% trim the bag and case the grains.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: Grey]
#21961311 - 07/18/15 04:40 PM (8 years, 6 months ago) |
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Yeah I also tought it could be a cube strain and not P. Mexicana, I will do that to see what kind of fruit grows from it, thanks guys.
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: My invitro techinque [Re: durxo]
#21961370 - 07/18/15 04:52 PM (8 years, 6 months ago) |
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Uh yeah that bag is extremely bacterial. That thick white look, and the wet grains.. i can't believe all you do is just use alcohol. Try using just alcohol on your scalpel for agar. I used to and it doesn't work. At all. Then I bought a blow torch. What a difference!
Oh BTW lab grade 99% alcohol is actually worse than 70% alcohol. Pure alcohol is nowhere near as good at sanitizing.
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durxo
Stranger



Registered: 01/16/14
Posts: 113
Last seen: 2 years, 1 month
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Re: My invitro techinque [Re: Mad Season]
#21961555 - 07/18/15 05:41 PM (8 years, 6 months ago) |
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Ok next time I will flame sterilize
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“The psychotic drowns in the same waters in which the mystic swims with delight.” ― Joseph Campbell
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