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OfflineSgt. Pepper
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Agar to lc idea
    #19405513 - 01/11/14 07:49 PM (10 years, 19 days ago)

So I have switched over from lc to agar since contams piss me off and agar really can give me plenty of inoculate. So I have an idea to get mycelium from an agar plate into a spawn bag. I don't have a flow hood so putting a wedge into the bag is unrealistic using a small SAB. I'm planning on making a lc-esque solution from the agar plate. I do agar in pp5 containers with a SHIP so I can put a drop of lc on the plate without opening it, it seems to be more sterile than opening it up even inside a SAB. So what I wanna do is inject sterile, distilled water into the pp5 containers, swirl it around a bit, and suck it bad up. This seems like a good idea since I can remain completely sterile and inoculate spawn bags without opening the bag. Has anyone done something like this? Is this a bad idea for any reason? So far I'm loving agar, I'm just worried it won't be practical for spawn bags without a flow hood which is just out of the question for me.


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OfflinecronicrFacebook
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405529 - 01/11/14 07:53 PM (10 years, 19 days ago)

yes i've done it, worked fine an no reason to use distilled, test it first:thumbup:


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Invisiblewildernessjunkie
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405552 - 01/11/14 07:58 PM (10 years, 19 days ago)

Its been done. Its even been done with PF cakes. Ive even done agar to LC successfully via FH.

If you have the SAB. And you can do agar. Then I would recommend G2G transfers instead.

I spent quite a bit of time fucking around with LC's. And for the most part have completely given them up. The speed and quantity of spawn that is possible via G2G is amazing. It quickly outpaces LC in every way.

Im not saying that LC isn't a good tool mind you. But having done both, I think that G2G is a significantly better choice.


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Invisiblebootster
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Re: Agar to lc idea [Re: wildernessjunkie]
    #19405561 - 01/11/14 08:01 PM (10 years, 19 days ago)

:whathesaid:


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OfflineSgt. Pepper
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Re: Agar to lc idea [Re: wildernessjunkie]
    #19405611 - 01/11/14 08:10 PM (10 years, 19 days ago)

Quote:

wildernessjunkie said:
Its been done. Its even been done with PF cakes. Ive even done agar to LC successfully via FH.

If you have the SAB. And you can do agar. Then I would recommend G2G transfers instead.

I spent quite a bit of time fucking around with LC's. And for the most part have completely given them up. The speed and quantity of spawn that is possible via G2G is amazing. It quickly outpaces LC in every way.

Im not saying that LC isn't a good tool mind you. But having done both, I think that G2G is a significantly better choice.



The only problem is that opening my spawn bags inside my SAB would be difficult and impractical, as well as a larger opportunity for a contaminant to ruin 5 pounds of birdseed. I wouldn't really say that I'm making a traditional lc either, I suppose I mis-spoke. On the shroomery, when someone thinks of lc, they picture a nutrient liquid, but I'm planning on using reverse osmosis water with nothing in it. After I get the mycelium suspended into the water I'll use it more or less immediately. G2G is an awesome method, I use to use it with quart jars, but sense I switched to spawn bags, I'm not sure how I could make it work.


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Invisiblewildernessjunkie
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405658 - 01/11/14 08:22 PM (10 years, 19 days ago)

What you are wanting to do has been done. And it has worked.

I do foresee some problems with getting the mycelium in suspension though. Some folks in the past have used a needle to scrape the myc off the agar. Others have taken the agar wedge and blended it with sterile water with an Oster brand blender.

Be aware that every step that you add along the way to inoculation, adds a contamination vector.


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InvisibleJ. Jack Flash
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405672 - 01/11/14 08:27 PM (10 years, 19 days ago)

i've sorta done this, albeit a much smaller scale.  there's a doc or two mentioning using dH2O for long term storage of mycelial cultures.  long term, like years long without refrigeration.  it's basically doing exactly this, swishing sterile water around a culture on agar, and storing it in dram vials.  i made a couple from multispore agar and noc'd a few jars to feel the process.  so far so good.  visible growth in just a day or three.

so as already stated, it works, and has the added benefit of usefulness for culture storage.  but i feel like g2g will be way faster if you can get away with it.


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OfflineGeneric
Registered: 11/12/13
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Re: Agar to lc idea [Re: J. Jack Flash]
    #19405680 - 01/11/14 08:29 PM (10 years, 19 days ago)

right...  back to the problem of using spawn bags in a still air box
:popcorn:


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OfflineSgt. Pepper
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Re: Agar to lc idea [Re: Generic]
    #19405695 - 01/11/14 08:32 PM (10 years, 19 days ago)

Quote:

wildernessjunkie said:
What you are wanting to do has been done. And it has worked.

I do foresee some problems with getting the mycelium in suspension though. Some folks in the past have used a needle to scrape the myc off the agar. Others have taken the agar wedge and blended it with sterile water with an Oster brand blender.

Be aware that every step that you add along the way to inoculation, adds a contamination vector.



Thank you, that's exactly what I wanted to hear. If it's been done successfully I'm happy, but typical 50/50 success rates with classic nutrient lc piss me off more than ever. I just restarted an old cubensis lc that was clean when I made it, but it had bacteria after just 3 months of storage. Plus you can't isolate from a lc! I'm still not sure why all the vendors sell edibles as lc's, I even got a trich syringe that was supposed to be blue oyster from a very reputable shroomery sponsor, if only they sold agar plates.
Quote:

J. Jack Flash said:
i've sorta done this, albeit a much smaller scale.  there's a doc or two mentioning using dH2O for long term storage of mycelial cultures.  long term, like years long without refrigeration.  it's basically doing exactly this, swishing sterile water around a culture on agar, and storing it in dram vials.  i made a couple from multispore agar and noc'd a few jars to feel the process.  so far so good.  visible growth in just a day or three.

so as already stated, it works, and has the added benefit of usefulness for culture storage.  but i feel like g2g will be way faster if you can get away with it.



Yeah, I've read this too and it seems like a really cool way to store a good isolate.
Quote:

Generic said:
right...  back to the problem of using spawn bags in a still air box
:popcorn:



I just don't think it's practical, maybe buy a really tall plastic tub. Lol.


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InvisibleJ. Jack Flash
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Re: Agar to lc idea [Re: Generic]
    #19405699 - 01/11/14 08:33 PM (10 years, 19 days ago)

place your still air box upside down on top of another tub with the same size opening and tape it shut... then don't drop anything inside it.  :laugh:


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2020 new years grow along


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OfflineSgt. Pepper
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Re: Agar to lc idea [Re: J. Jack Flash]
    #19405705 - 01/11/14 08:36 PM (10 years, 19 days ago)

Quote:

J. Jack Flash said:
place your still air box upside down on top of another tub with the same size opening and tape it shut... then don't drop anything inside it.  :laugh:



The arm holes would have to be arm ovals. Haha. Or whatever shape a track is.


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InvisibleJ. Jack Flash
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405724 - 01/11/14 08:41 PM (10 years, 19 days ago)

wah wah.  didn't say it was a great idea...  :smile:


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OfflineSgt. Pepper
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Re: Agar to lc idea [Re: J. Jack Flash]
    #19405736 - 01/11/14 08:43 PM (10 years, 19 days ago)

Quote:

J. Jack Flash said:
wah wah.  didn't say it was a great idea...  :smile:



Lol, I do really wish I could put a spawn bag inside a box, that would open up a world of opportunity.


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Offlinekaptanoblivious
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Registered: 11/24/13
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405772 - 01/11/14 08:52 PM (10 years, 19 days ago)

Quote:

Sgt. Pepper said:
Plus you can't isolate from a lc!




Wait, why can't you isolate from an LC? Just put a drop on an agar plate, and streak it out a few times with a sterile loop, using the standard streak plate method. Is this not going to work?


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OfflinecronicrFacebook
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Re: Agar to lc idea [Re: kaptanoblivious]
    #19405779 - 01/11/14 08:53 PM (10 years, 19 days ago)

well then it's agar your isolating with......


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It doesn't matter what i think of you...all that matters is clean spawn

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Invisiblewildernessjunkie
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Re: Agar to lc idea [Re: cronicr]
    #19405808 - 01/11/14 08:58 PM (10 years, 19 days ago)

Quote:

cronicr said:
well then it's agar your isolating with......




Yep.

And as such, would just be easier to start that way in the first place. And then you wouldn't take the chance of isolating contamination.


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OfflinecronicrFacebook
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Re: Agar to lc idea [Re: wildernessjunkie]
    #19405810 - 01/11/14 08:59 PM (10 years, 19 days ago)

it's true:thumbup:


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It doesn't matter what i think of you...all that matters is clean spawn

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Offlinekaptanoblivious
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Re: Agar to lc idea [Re: cronicr]
    #19405826 - 01/11/14 09:04 PM (10 years, 19 days ago)

I thought he meant you couldn't get an isolate once it went to liquid culture. Nothing to see here, move along.
:irdumbtard:


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InvisibleJ. Jack Flash
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Re: Agar to lc idea [Re: Sgt. Pepper]
    #19405832 - 01/11/14 09:06 PM (10 years, 19 days ago)

all right, sarge, now you got me thinkin. 
:hmm:
what's really stopping it?  the height of the open bag, right?  hmmm... also the number of bags you could fit in the box.  turn a tub on its side?  arm slits in a garment bag?  custom positive pressure hepa filtered cardboard box?  robot arms in a vacuum chamber?  mice?

frank is apparently planning a write up on the subject.  so it must be possible.


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OfflineSgt. Pepper
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Re: Agar to lc idea [Re: J. Jack Flash]
    #19405884 - 01/11/14 09:19 PM (10 years, 19 days ago)

Quote:

J. Jack Flash said:
all right, sarge, now you got me thinkin. 
:hmm:
what's really stopping it?  the height of the open bag, right?  hmmm... also the number of bags you could fit in the box.  turn a tub on its side?  arm slits in a garment bag?  custom positive pressure hepa filtered cardboard box?  robot arms in a vacuum chamber?  mice?

frank is apparently planning a write up on the subject.  so it must be possible.



I use a SAB for inoculating bags with a syringe, but the tops are pushed up against the lid so I couldn't do grain to grain with them since the lid is in the way. To be honest, only a couple more inches would be necessary. It's probably doable, it just seems difficult and contaminate prone. There's something I just love about the sterile security of a syringe though. For example, I'd prefer to do a glc than a g2g transfer.


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