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OfflineLoveLightPeace
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Question about an inoculation procedure
    #18725684 - 08/19/13 03:11 AM (1 year, 3 months ago)

I read the following in a mushroom grow book as being one of the most sterile procedures for inoculation and wud like some feedback:

"This method is carried out as follows: Sterilize a bit of water by placing it in a flask sealed with aluminum foil and placing in the PC with the mason jars. When one is ready to inoculate, take up about 10 cc (ml) of the water in a disposable, presterilized plastic syringe, and inject it onto the surface of an agar mycelial culture (slant-tube cultures are ideal for this). Reseal the culture and swirl the water around vigorously until fragments of mycelium become visible suspended in the water. Then again take up the water-mycelial suspension in the syringe, and use the syringe to inoculate
each mason jar by injecting 1 or 2 ccs into each jar."
(Taken from "psilocybin: magic mushroom grower's guide pg. 40)


My Qs:

1-Is this a good way to inoculate or is it sufficient to just simply drop a wedge into the spawn jar from a petri dish?

2-i know agar is an inoculant/inoculum and grains are used as substrate/spawn, but is agar/malt extract generally less prone to contams than say WBS?


--------------------
I am so small I can barely be seen.
How can this great love be inside me?

Look at your eyes. They are small,
but they see enormous things.

(Rumi)


Edited by LoveLightPeace (08/19/13 03:19 AM)


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: LoveLightPeace]
    #18725723 - 08/19/13 03:27 AM (1 year, 3 months ago)

Hm, never heard of this before, but now I'm thinking of using it. I love liquid culture.

Lets see what the pros think of it though :hatsoff:


--------------------
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Offlinevan der griegen
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Re: Question about an inoculation procedure [Re: LoveLightPeace]
    #18725736 - 08/19/13 03:35 AM (1 year, 3 months ago)

Quote:

LoveLightPeace said:
I read the following in a mushroom grow book as being one of the most sterile procedures for inoculation and wud like some feedback:

"This method is carried out as follows: Sterilize a bit of water by placing it in a flask sealed with aluminum foil and placing in the PC with the mason jars. When one is ready to inoculate, take up about 10 cc (ml) of the water in a disposable, presterilized plastic syringe, and inject it onto the surface of an agar mycelial culture (slant-tube cultures are ideal for this). Reseal the culture and swirl the water around vigorously until fragments of mycelium become visible suspended in the water. Then again take up the water-mycelial suspension in the syringe, and use the syringe to inoculate
each mason jar by injecting 1 or 2 ccs into each jar."
(Taken from "psilocybin: magic mushroom grower's guide pg. 40)


My Qs:

1-Is this a good way to inoculate or is it sufficient to just simply drop a wedge into the spawn jar from a petri dish?

2-i know agar is an inoculant/inoculum and grains are used as substrate/spawn, but is agar/malt extract generally less prone to contams than say WBS?




1.  Dropping a wedge into a master jar is sufficient.  The quoted text has some problems, aside from being needlessly complicated for someone who already has a clean culture on agar, as well.  Be very careful when sterilizing empty syringes.  If you have the plunger depressed, the inside of the syringe presents a good entry point for contamination until the plunger is drawn back.

2.  No, agar/malt is no more or less prone to contamination than grains.  Malt is just the dried extract of a sprouted grain that's had its germination halted by heat.  Agar is superior for dealing with the contaminations, however, since you have what's essentially a two-dimensional surface with clearly visible contaminations which can be transferred away from.


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OfflineBloodKil
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Re: Question about an inoculation procedure [Re: van der griegen]
    #18725744 - 08/19/13 03:38 AM (1 year, 3 months ago)

:whathesaid:


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: BloodKil]
    #18725746 - 08/19/13 03:40 AM (1 year, 3 months ago)

thats a good point, but wouldn't this be a good way to make a big LC jar from an isolate?


--------------------
Everything's blue in this world, the deepest shade of mushroom blue. NIN


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Offlinevan der griegen
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726557 - 08/19/13 10:50 AM (1 year, 3 months ago)

Why would you want to make an LC when you can use G2G?


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: van der griegen]
    #18726590 - 08/19/13 10:59 AM (1 year, 3 months ago)

Quote:

van der griegen said:
Why would you want to make an LC when you can use G2G?



? Does it have to be one or the other? I like the ease of a syringe.

And with 4 little squirts of solution, you have 4 innoc points. Starts faster than G2G does for me.


--------------------
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OfflineMush4Brains
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726596 - 08/19/13 11:01 AM (1 year, 3 months ago)

Quote:

rbalzer said:
thats a good point, but wouldn't this be a good way to make a big LC jar from an isolate?




If you want a water LC take a wedge, put it into a half pint filled up 1/3rd full with grain, let that colonize, then make a grain LC from that.  No reason to fuck with a nice culture on agar more than you have to.


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: Mush4Brains]
    #18726617 - 08/19/13 11:06 AM (1 year, 3 months ago)

Quote:

Mush4Brains said:
Quote:

rbalzer said:
thats a good point, but wouldn't this be a good way to make a big LC jar from an isolate?




If you want a water LC take a wedge, put it into a half pint filled up 1/3rd full with grain, let that colonize, then make a grain LC from that.  No reason to fuck with a nice culture on agar more than you have to.



Um, that doesn't make sense dude. That only adds an extra step.

In what way is it better to make LC with grains than wedge? I've got news for you noob, its not :shrug:


--------------------
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Offlinevan der griegen
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726622 - 08/19/13 11:07 AM (1 year, 3 months ago)

Quote:

rbalzer said:
Quote:

van der griegen said:
Why would you want to make an LC when you can use G2G?



? Does it have to be one or the other? I like the ease of a syringe.

And with 4 little squirts of solution, you have 4 innoc points. Starts faster than G2G does for me.




Really?  It's always been my experience that LC sulks for two or three days before finally showing up for the party, while G2G is off like a horse at the races.  I usually see it recover in less than a day, and the reasoning is pretty sound:  it's already got its hyphae into and consolidated a pretty good source of both nutrients and water, whereas a piece of mycelium from an LC has to do that before it can spread in earnest.

And as far is inoculation points go, once I shake my jars up I have dozens.  That's way better than four.


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: van der griegen]
    #18726630 - 08/19/13 11:08 AM (1 year, 3 months ago)

Quote:

van der griegen said:
Quote:

rbalzer said:
Quote:

van der griegen said:
Why would you want to make an LC when you can use G2G?



? Does it have to be one or the other? I like the ease of a syringe.

And with 4 little squirts of solution, you have 4 innoc points. Starts faster than G2G does for me.




Really?  It's always been my experience that LC sulks for two or three days before finally showing up for the party, while G2G is off like a horse at the races.  I usually see it recover in less than a day, and the reasoning is pretty sound:  it's already got its hyphae into and consolidated a pretty good source of both nutrients and water, whereas a piece of mycelium from an LC has to do that before it can spread in earnest.



well maybe I need a little more experience. I've only done LC inoculations a few times, but I thought I remembered it being quick. Like as fast as G2G. Just because its in water shouldn't make it take an extra few days, I'd think :shrug:


--------------------
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OfflineMush4Brains
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726640 - 08/19/13 11:11 AM (1 year, 3 months ago)

Quote:

rbalzer said:
Quote:

Mush4Brains said:
Quote:

rbalzer said:
thats a good point, but wouldn't this be a good way to make a big LC jar from an isolate?




If you want a water LC take a wedge, put it into a half pint filled up 1/3rd full with grain, let that colonize, then make a grain LC from that.  No reason to fuck with a nice culture on agar more than you have to.



Um, that doesn't make sense dude. That only adds an extra step.

In what way is it better to make LC with grains than wedge? I've got news for you noob, its not :shrug:




What are talking about?  I'm sure if I wanted to make an LME LC or something of the sort, you could drop a wedge right into it, sure, but mycelium water is probably the best LC I've ever used.  If OP doesn't want to fuck with an agar plate with a nice iso on it, what better way would there be to make a grain LC than what I have said?

And you think that because my registration date is new somehow I'm a "noob?"  Come on now.


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OfflinePussyFartM
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726646 - 08/19/13 11:13 AM (1 year, 3 months ago)

The fact that you have to wait for the LC to colonize means it is slower than G2G...

You are going to inoculate grains with the LC anyways right?

It's basically just an extra, unneeded step that only wastes time and adds contamination risk if you ask me.

Just do spores or culture to grains, and then G2G....it is the best and fastest way to expand your spawn.


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: Mush4Brains]
    #18726647 - 08/19/13 11:13 AM (1 year, 3 months ago)

Quote:

Mush4Brains said:
Quote:

rbalzer said:
Quote:

Mush4Brains said:
Quote:

rbalzer said:
thats a good point, but wouldn't this be a good way to make a big LC jar from an isolate?




If you want a water LC take a wedge, put it into a half pint filled up 1/3rd full with grain, let that colonize, then make a grain LC from that.  No reason to fuck with a nice culture on agar more than you have to.



Um, that doesn't make sense dude. That only adds an extra step.

In what way is it better to make LC with grains than wedge? I've got news for you noob, its not :shrug:




What are talking about?

Mycelium water is probably the best LC I've ever used. 

And you think that because my registration date is new somehow I'm a "noob?"  Come on now.



what are YOU talking about. You literally make no sense. I feel I logically stated why i think you're wrong. And you need to tell me WHY i'm wrong.

And its logical to assume you're a noob, since you registered this year and don't appear to know what you're talking about :lol:


--------------------
Everything's blue in this world, the deepest shade of mushroom blue. NIN


:aliendance::awedance:TRADE LIST:awedance::aliendance:


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: PussyFart]
    #18726652 - 08/19/13 11:15 AM (1 year, 3 months ago)

Quote:

Notahacker420 said:
The fact that you have to wait for the LC to colonize means it is slower than G2G...

You are going to inoculate grains with the LC anyways right?

It's basically just an extra, unneeded step that only wastes time and adds contamination risk if you ask me.

Just do spores or culture to grains, and then G2G....it is the best and fastest way to expand your spawn.



Thats a good point. I suppose the only advantage is not having to open all the jars momentarily to G2G. I have a bad contam rate, maybe thats why I like syringe LC.

thanks for explaining it to me everyone.


--------------------
Everything's blue in this world, the deepest shade of mushroom blue. NIN


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OfflineMush4Brains
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Re: Question about an inoculation procedure [Re: rbalzer]
    #18726660 - 08/19/13 11:20 AM (1 year, 3 months ago)

Quote:

rbalzer said:
what are YOU talking about. You literally make no sense. I feel I logically stated why i think you're wrong. And you need to tell me WHY i'm wrong.

And its logical to assume you're a noob, since you registered this year and don't appear to know what you're talking about :lol:




What do you mean I made no sense?  That is how you start with agar and make a grain liquid culture, also called a mycelium water, or a mycelium slurry.  Which is specificially what the OP was asking about in his post, inoculating with a water culture.  He asked if he should shoot water all over a petri, I don't agree that this would be the best way to go about it, so I suggest expanding to grains first.

You take a wedge, use it to inoculate a small amount of grains, shoot it up with water and shake it around, and then suck your sugar-free LC back into the syringe?  What don't you get about this? 


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Offlinerbalzer
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Re: Question about an inoculation procedure [Re: Mush4Brains]
    #18726763 - 08/19/13 11:49 AM (1 year, 3 months ago)

Quote:

Mush4Brains said:
Quote:

rbalzer said:
what are YOU talking about. You literally make no sense. I feel I logically stated why i think you're wrong. And you need to tell me WHY i'm wrong.

And its logical to assume you're a noob, since you registered this year and don't appear to know what you're talking about :lol:




What do you mean I made no sense?  That is how you start with agar and make a grain liquid culture, also called a mycelium water, or a mycelium slurry.  Which is specificially what the OP was asking about in his post, inoculating with a water culture.  He asked if he should shoot water all over a petri, I don't agree that this would be the best way to go about it, so I suggest expanding to grains first.

You take a wedge, use it to inoculate a small amount of grains, shoot it up with water and shake it around, and then suck your sugar-free LC back into the syringe?  What don't you get about this? 



thats what I wanted. You to elaborate, aka explain it better. that makes more sense.

Sorry for bein a dick. with your first couple posts you really sounded like a noob to me, obviously :shrug:

:douchewink:


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