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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
vacum pulled spores into lc...
    #7721557 - 12/05/07 07:53 PM (16 years, 1 month ago)

so instead of putting the 2 cc's of spore solution into the 400 ml's of lc mix the vacum pulled between 5-6 cc's in two of the jars i knocked up. is this going to be okay? what harm could come of this?


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OfflineThe Shape

Registered: 10/27/07
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7721603 - 12/05/07 08:01 PM (16 years, 1 month ago)

as long as the solution wasnt too diluted the mycelium should drink up any excess water

a few extra CCs shouldnt hurt as long as there isnt a large puddle covering the bottom of the jar


--------------------
You miserable, presumptuous, mediocre piece of shit. You have no talent. Your no artist. An artist respects the silence, it serves as the foundation of creativity. Your a disgrace. Your everything thats gone wrong in this world. You're self consumed, no talent, mediocre piece of shit. And i've earned my right to say it.  In 1975 i walked Bob Dylan up on stage. Who the fuck are you?
I knew The Grateful Dead in 1966. Who the fuck are you?
You're nothing... And you'll never be anything.

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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
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Re: vacum pulled spores into lc... [Re: The Shape]
    #7721652 - 12/05/07 08:10 PM (16 years, 1 month ago)

Quote:

NIBRAL said:
as long as the solution wasnt too diluted the mycelium should drink up any excess water

a few extra CCs shouldnt hurt as long as there isnt a large puddle covering the bottom of the jar





i guess im not following. how could their be a puddle at the bottom of the jar if its all liquid?


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OfflineBlargIAmDead
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7721686 - 12/05/07 08:15 PM (16 years, 1 month ago)

Nibral didn't read thoroughly:). And it should be fine. You can OVER innoc an LC. It'll go to the point of using all the sugars and then stop.


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OfflineThe Shape

Registered: 10/27/07
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Re: vacum pulled spores into lc... [Re: BlargIAmDead]
    #7721765 - 12/05/07 08:29 PM (16 years, 1 month ago)

my fault i ddint see LC


--------------------
You miserable, presumptuous, mediocre piece of shit. You have no talent. Your no artist. An artist respects the silence, it serves as the foundation of creativity. Your a disgrace. Your everything thats gone wrong in this world. You're self consumed, no talent, mediocre piece of shit. And i've earned my right to say it.  In 1975 i walked Bob Dylan up on stage. Who the fuck are you?
I knew The Grateful Dead in 1966. Who the fuck are you?
You're nothing... And you'll never be anything.

- Douglas Levison


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Invisibletakk
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Re: vacum pulled spores into lc... [Re: The Shape]
    #7722086 - 12/05/07 09:48 PM (16 years, 1 month ago)

in the future you can equalize the vacuum in a jar by inserting a syringe with the plunger taken out and stuffed with poly-fill to ensure a minimum of airborne contaminants enter the jar, then proceed to inoculate with your other syringe (with the correct amount of cc's you desire)

:stoned:


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Invisiblemonstermitch
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Re: vacum pulled spores into lc... [Re: takk]
    #7722102 - 12/05/07 09:53 PM (16 years, 1 month ago)

well there never should have been a vacuum to begin with.

Liquid Culture needs gas exchange.
a vacuum means that there is no gas exchange.
so if you bought that from somebody, they jacked you, sorry.

an LC with no gas exchange will only partially colonize, then it
will stall out.

you have the option of making your own airport out of a needle
and some poly-fil, but why would you want to take a chance like
that?
if you leave the airport in for too long, it'll ruin the injection site
and leave a permanent hole in it...

all lids for liquid culture need some sort of filter material built into
them to provide correct gas exchange. otherwise you're asking
for trouble, more trouble than just a vacuum and over-inoculation.


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



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Invisibletakk
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Re: vacum pulled spores into lc... [Re: monstermitch]
    #7722119 - 12/05/07 09:56 PM (16 years, 1 month ago)

didn't thinkathat :wink:


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OfflineSeventy
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Re: vacum pulled spores into lc... [Re: takk]
    #7722206 - 12/05/07 10:22 PM (16 years, 1 month ago)

an LC doesn't need that much gas exchange, barely any at all imho. the myc is suspended in liquid, honestly how much gas is getting to it? there isn't a bubbler stone in there keeping the gas up, so whatever O2 is in there will get sucked up quick. it's only going to colonize how much nutrients are in there, and eventually it will stall out anyways

having it vacuumed is just plain wrong though


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Invisiblemonstermitch
Growing in Bags Doesn't Work

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Re: vacum pulled spores into lc... [Re: Seventy]
    #7722323 - 12/05/07 10:42 PM (16 years, 1 month ago)

well is your honest opinion based on experience or just a guess?

I'm guessing it's the latter.

my opinion is based 100% on experience of trial and error.
on side by side tests.

there's plenty of proof here on this site to support my claim as
well, not just my own personal experience.

so I guess we'll just have to agree to disagree.

by the way, an air stone or bubble stone would provide air
exchange. which has nothing to do with colonizing media.
we're talking gas exchange here, which is completely different
from air exchange, which is for fruiting substrates.


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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: monstermitch]
    #7723223 - 12/06/07 06:20 AM (16 years, 1 month ago)

no imade the lids myself, and the medium. i followed a tek which said nothing about gas-exchange.

is there anyway i could somehow add something at this point to give it gas exchange?

how did this become a vacum in the first place? should i have put the lids rubber side up? not tightened them so much?


MM- where do you get lids that both have an injection port and something for gas exchange?


Edited by blood4blood (12/06/07 06:21 AM)


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Offlinekillerthc
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7723531 - 12/06/07 08:26 AM (16 years, 1 month ago)

just get a filter disc and some silicone and put 2 holes in a jar lid and fill one with silicone and put the filter disc over the other.


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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: killerthc]
    #7723563 - 12/06/07 08:38 AM (16 years, 1 month ago)

Quote:

killerthc said:
just get a filter disc and some silicone and put 2 holes in a jar lid and fill one with silicone and put the filter disc over the other.






yeah but is this something that i can do now? i need to get some sort of gas exchange in there now because they are under a vacum i think.


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OfflineRogerRabbitM
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7723593 - 12/06/07 08:49 AM (16 years, 1 month ago)

The more air to a liquid culture the better. That's why experienced growers always use a magnetic stirrer running 24/7. This ensures the entire LC is constantly oxygenated. I still don't understand why growers are slowing down their work so much trying to grow liquid cultures in mason jars. You can expand mycelium at least ten times faster in grains than in liquid, AND you'll know if it's contaminated before you use it.
RR


--------------------
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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."
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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: RogerRabbit]
    #7724754 - 12/06/07 02:23 PM (16 years, 1 month ago)

would it be possible under complete sterile conditions to puncture the lid with a sterile nail in a glovebox and quick put a piece of micropore tape over that hole for gas exchange?


RR- are you referring to G2G transfers???


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Offlinekillerthc
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7724775 - 12/06/07 02:28 PM (16 years, 1 month ago)

i think what you could do is sterilize a new lid after putting the holes in it then using a glove box replace the lid of your LC.


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OfflineRogerRabbitM
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Re: vacum pulled spores into lc... [Re: killerthc]
    #7724857 - 12/06/07 02:58 PM (16 years, 1 month ago)

Quote:

RR- are you referring to G2G transfers???




Sort of, but if you want liquid culture syringes, all you have to do is grow out a quart jar of rye berries, and then when it's fully colonized, shake it well, and pour nearly a full quart of sterilized water into the jar. Shake well again, and then draw the myceliated water back out into syringes. As an alternative, you can pour the quart of myceliated water into a sterile gallon of water to dilute the solution. You'll then have enough liquid culture to fill a few hundred syringes, and it only takes two to three weeks to get to this point from spores.

In addition, after harvesting the mycelium from the surface of the grains as described above, you can then still use the left over grains to inoculate ten to twenty additional jars of grains via grain to grain transfer in the traditional method.
RR


--------------------
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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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OfflineNibin
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Re: vacum pulled spores into lc... [Re: RogerRabbit]
    #7724900 - 12/06/07 03:07 PM (16 years, 1 month ago)

can this be done with a PF cake Roger?

I can imagine you can add less water because of the consistency, but can you get enough for a pint of LC?

EDIT: Or will the dry verm barrier be an issue?


--------------------
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Edited by Nibin (12/06/07 03:09 PM)


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Invisibleblood4blood
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Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: Nibin]
    #7724926 - 12/06/07 03:12 PM (16 years, 1 month ago)

awesome... ijust pc'd 10 qrts of rye and was thinking about doing agars grain lc tek once they colonized. i think thats the same thing you are talking about.


very simple, thanks a million for the info RR


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OfflineRogerRabbitM
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7724952 - 12/06/07 03:19 PM (16 years, 1 month ago)

Yea, agar wrote that up after reading a few of my posts about it. I believe fahtster also came up with the idea. For pf cakes, just skip the verm barrier. It's obsolete anyway now that we have micropore tape. Just tape up the inoculation holes before you sterilize, and then after inoculation, put a fresh piece of tape over the needle damaged tape.
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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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: RogerRabbit]
    #7724971 - 12/06/07 03:23 PM (16 years, 1 month ago)

RR- what do you think of my suggestion of piercing a hole in the lids of my 3 lc jars in a glove box for a ge port? or will they be okay with out one. there is 400 ml of medium in a quart.

under sterile conditions of course..

would it suck in a bunch of air being that its probabally under a vacum?


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OfflineRogerRabbitM
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7725119 - 12/06/07 03:57 PM (16 years, 1 month ago)

Wrap an alcohol soaked paper towel around the nail when you pierce the lid. The towel will act as a filter.
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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Invisibleblood4blood
Calmer Than You Are


Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: vacum pulled spores into lc... [Re: RogerRabbit]
    #7729112 - 12/07/07 02:55 PM (16 years, 1 month ago)

so i just decided to bump this again instead of starting a new thread.


so i went ahead and took RR's advice and used a alcohol soaked paper towel and piereced my lc jars. (good thing i did this because i heard air whoosing in as soon as i pierced the lid)

my question is- when i piereced the lid with the nail i noticed that a few drops of alcohol dripped down the nail and into the lc medium. will this be a problem? will it kill the spores?


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OfflineFraggin
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Re: vacum pulled spores into lc... [Re: blood4blood]
    #7729124 - 12/07/07 02:57 PM (16 years, 1 month ago)

A few drops? I say no.


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Invisiblemonstermitch
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Re: vacum pulled spores into lc... [Re: Fraggin]
    #7729278 - 12/07/07 03:34 PM (16 years, 1 month ago)

I agree, that shouldn't be too much trouble.

be careful to never get the micropore tape you put over those
holes wet.
if you do, expect certain contamination.

wet micropore tape serves as a wick to introduce contams into
the jar, which is no good.

getting it wet with the sugar-laden LC liquid is in even worse idea.

so just be careful and keep that micropore dry.


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OfflineBlargIAmDead
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Re: vacum pulled spores into lc... [Re: monstermitch]
    #7729363 - 12/07/07 03:59 PM (16 years, 1 month ago)

Okay, I have a question. I tape my jars before I foil wrap and boil them. It is inevitable the tape gets wet. What would be the proper procedure for sterility's sake? Should I leave them in the pot, with the foil covered till I'm ready to inoculate, or can I take them out of the pot, remove the foil, and put them (still pretty warm) into a still air box to cool off since there's a dry verm barrier? Or should I just leave the tape off period till after I inoculate?


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Invisibleblood4blood
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Re: vacum pulled spores into lc... [Re: BlargIAmDead]
    #7729579 - 12/07/07 04:39 PM (16 years, 1 month ago)

if there's a dry verm layer then there is no need for the tape.


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