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Psilosopherr
A psilly goose



Registered: 02/15/12
Posts: 12,278
Last seen: 1 month, 10 days
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One way valve to preserve sterility of invitro grows when adding water
#22302098 - 09/28/15 10:22 AM (8 years, 4 months ago) |
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I've been liking the idea of invitro grows lately because you can get much cleaner spore prints/clone tissue. But most people who do this give up on sterility as soon as the substrate needs water, they just open the jar lid and pour water right inside.
The 3 options: I'm thinking it might work to make some custom lids with one way valves installed. Either this or just have a SHIP and fill a large syringe with sterile water and water it that way. Or really I could just open the jar in a SAB and water it then.
Somehow a one way valve seems like a better idea to me. What are your thoughts?
One potential problem is pouring water into the valve. Will dirty air mix in with the sterile water as it pours? I don't think so, but That and who knows if they make autoclavable valves
Idk, what seems like the better method of sterile watering and why?
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22302592 - 09/28/15 12:26 PM (8 years, 4 months ago) |
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The sterile water and the SHIP seems about as good as it's going to get..
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Inocuole]
#22302615 - 09/28/15 12:33 PM (8 years, 4 months ago) |
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most subs should have enough water for a good flush without having to add any water
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Psilosopherr
A psilly goose



Registered: 02/15/12
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: cronicr]
#22302627 - 09/28/15 12:37 PM (8 years, 4 months ago) |
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Quote:
cronicr said: most subs should have enough water for a good flush without having to add any water
right. I was just wanting to try the bottom watering + grass seed method from one of violets teks.
Also I'm doing a little experiment where I use only a whole SFD with the band part of a metal lid. So I figure it will need the extra water with all that extra air getting through.
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22302651 - 09/28/15 12:43 PM (8 years, 4 months ago) |
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i see i see i've always just done typical cakes...seemed easier good luck blazer
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Psilosopherr
A psilly goose



Registered: 02/15/12
Posts: 12,278
Last seen: 1 month, 10 days
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: cronicr]
#22302668 - 09/28/15 12:48 PM (8 years, 4 months ago) |
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Quote:
cronicr said: i see i see i've always just done typical cakes...seemed easier good luck blazer
sure has been a long time since I've messed around with good ol' pf cakes.
This experiment just got a little more interesting now that cron himself can't think of a definite reason why it wouldn't work off the top of his head.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22302786 - 09/28/15 01:05 PM (8 years, 4 months ago) |
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Sterile invitro teks are here... Bulk growers best stay clear... The way of the future is near

brings me back to the halonium days
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: PirateSwazey]
#22302862 - 09/28/15 01:20 PM (8 years, 4 months ago) |
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I think if you're that worried about keeping things sterile you need to work on other things 
unless it's pure novelty.
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Psilosopherr
A psilly goose



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Re: One way valve to preserve sterility of invitro grows when adding water [Re: bodhisatta]
#22302872 - 09/28/15 01:22 PM (8 years, 4 months ago) |
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Quote:
bodhisatta said: I think if you're that worried about keeping things sterile you need to work on other things 
unless it's pure novelty.
other things?
Nothing wrong with cleaner spore prints. Sterile fungus flesh would be nice for making LC's too.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22302913 - 09/28/15 01:31 PM (8 years, 4 months ago) |
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the trouble it takes to get sterile mushroom flesh for an LC, you can make a petri dish, besides 9 times out of 10, or more than that my tissue biopsy to a dish is 100% clean, so in theory you could biopsy to a LC if you wanted, but I wouldn't.
as for spore prints, they're clean enough if you do indoor in a fruiting chamber let alone in a in-vitro setup. let alone trying to water them aseptic. seems like a moot concern, like being worried about the paint job on a car that's missing wheels.
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Psilosopherr
A psilly goose



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Re: One way valve to preserve sterility of invitro grows when adding water [Re: bodhisatta]
#22302957 - 09/28/15 01:42 PM (8 years, 4 months ago) |
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Alright I see what you mean.
Not enough extra work to worry about IMO though. Since I'm doing invitro regardless.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22302977 - 09/28/15 01:47 PM (8 years, 4 months ago) |
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you could add an inline 0.2 micron filter and inject water thru that into the container. I think they make syringe filters that will sterile filter liquids.
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Psilosopherr
A psilly goose



Registered: 02/15/12
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: bodhisatta]
#22303002 - 09/28/15 01:53 PM (8 years, 4 months ago) |
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Quote:
bodhisatta said: you could add an inline 0.2 micron filter and inject water thru that into the container. I think they make syringe filters that will sterile filter liquids.
you know I actually might have a few of those. I got em in a trade and didn't really know what they were for.
many thanks
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Psilosopherr
A psilly goose



Registered: 02/15/12
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Last seen: 1 month, 10 days
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22309838 - 09/29/15 05:34 PM (8 years, 3 months ago) |
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The filters I have seem like they'll work. I just hope the filter won't be compromised from all the water flow.
Maybe I'll silicone it right into the SFD I've been using as a lid.
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bodhisatta 
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Registered: 04/30/13
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Re: One way valve to preserve sterility of invitro grows when adding water [Re: Psilosopherr]
#22309877 - 09/29/15 05:41 PM (8 years, 3 months ago) |
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If it's .2 micron its made to filter liquid.
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mushpunx
Fungus Punk



Registered: 04/20/14
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Quote:
rbalzer said:
Quote:
bodhisatta said: I think if you're that worried about keeping things sterile you need to work on other things 
unless it's pure novelty.
other things?
Nothing wrong with cleaner spore prints. Sterile fungus flesh would be nice for making LC's too.
I dunno man I mean I suppose if you are making spore syringes than yea it would be nice to have an invitro cap to print with, provided your sterile tech is up to par..
But even the dirtiest print can be cleaned up with a few transfers on agar, and for cloning to LC you don't need the whole fruit to be sterile you just tear it open and biopsy a piece of virgin tissue... but again I would put this to agar too.
Dont skip the agar man, its the best tool we have
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 Amateur Mycologists United AMU Q&A
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
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I've never had problems with my prints from SGFC, or open air, humidity tent fruited grows. and no one I've sent a print too has had a problem either, swabs too. they've made their way into many syringes and no problems there either. the worry is null and void if you're clean when making the print. if you're paranoid print the cap for an hour then move it to a new piece of foil. the idea being that any contaminants would be deposited and when you make the second print that print will be much more clean.
you're getting more than billions of spores on the print and maybe a few viable contaminants which get diluted way down or not even grabbed depending on how much print you use. prints are 99,99% clean if you do them right
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mushpunx
Fungus Punk



Registered: 04/20/14
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Last seen: 11 days, 10 hours
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Yea man the way I see it is the only thing you will use a spore syringe for is PF cakes
I have never ever lost a PF cake to contamination.. even when I was just learning how to make syringes (and it was realll sloppy).. they seem pretty forgiving in that aspect
If a print from an indoor fruit is so dirty that you cant get clean growth from it on agar then somebody probably fucked up somewhere real bad, its not gunna be because the cap was dirty
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 Amateur Mycologists United AMU Q&A
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Psilosopherr
A psilly goose



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the preliminary results on SFD only lids were pleasing. I have two quart jars each knocked with the same isolate. One with a normal loose lid and the other with an SFD lid. They pinned around the same time I think. But the SFD lid shrooms are getting bigger faster. And they don't look all unhappy like most invitro fruits, they look like they actually have enough air.
bottom watering seems to do the trick.
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