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Offlinelgm0420
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Registered: 02/18/09
Posts: 134
Last seen: 11 years, 7 months
Solve this problem, earn a cookie...
    #18398729 - 06/10/13 06:26 PM (11 years, 7 months ago)

Hey everyone,

Im a long time reader on the forums, short time contributer information wise.  I have a problem which just wont get solved.  And yes, i could just move on from it since its not working for me, and I have proven methods to get results, but as a person who enjoys the scientific process learning through trial and error, i cant let this go.  Let me give you the short of it...

So I have been a strong follower of grain to grain transfers, its my bread and butter and what works best for me, but i recently decided i wanted to take on liquid cultures to see if it could help me in spawning grain faster and inoculating grain bags.  I had some success with my first turbinado/water LC.  But it was limited to this one particular liquid culture, and i had no success after.

Once i hit the wall, i went at it trying to figure out what was going on.  I had some contams, so i thought bad LC.  Made new LCs.  Now the problem wasn't bacterial growth, it was growth of any kind.  So i tried different volumes of LC on inoculation.  I had bags with 4 quarts of grain, so i tried anything from 12 to 25 mLs a bag.  Only growth i got was at the higher dose of LC, but it was bacterial and took at least 2 weeks to set in.  Most of the bags showed no growth at all, ever.  Im talking, sittin in the incubator for 3 to 4 weeks just to see if it'll grow anything.  So i thought water content.  I simmered for different times, soaked for different times, didnt simmer certain batches.  Tried using coffee to help pH.  Nothing helped the problem.  So i thought maybe it was the medium that the LC was growing on.  Left sugar in the raw (turbinado) and went and tried malt extract, and malt/dex combo.  LCs looked great but nothing, no growth yet again.  This last one has me clueless.  I dont know where to go from here.

All of these LCs are spawned from clean grain LCs, some from the same grain LCs some from different.  All looked robust and healthy, but none would transfer over to the grain.  Incubated at a steady 78 degree incubator that proves to work with everything else i put in there.  Started soaking grain for 24 hours, then simmered for half hour, cooled and PCd.  That is my normal protocol, but when that failed to work, i branched out.  I was thinking maybe the grain is no good?  but its the same grain i have bought from whole foods for years.  Rye berries.

Anyone have ideas as to what i got goin wrong here? im stumped, and i dont like it.

thanks!

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Offlinezpores
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Registered: 11/30/10
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Re: Solve this problem, earn a cookie... [Re: lgm0420]
    #18399006 - 06/10/13 07:37 PM (11 years, 7 months ago)

How are you getting your grain water? The water isn't boiling hot when you shoot it into your grain is it? lol sounds dumb but you didn't really mention your technique.

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OfflineJii
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Registered: 01/17/08
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Re: Solve this problem, earn a cookie... [Re: lgm0420]
    #18399706 - 06/10/13 09:29 PM (11 years, 7 months ago)

Why don't you use your clean GLC straight to bags?
That's what I do and it works like a charm.
I just inoculate small rye jar, wait it to colonise, shake it loose,
fill with water, gentle shake again, then fill bunch of syringes.

It doesn't take too much GLC to start one bag. :stoned:


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Offlinelgm0420
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Registered: 02/18/09
Posts: 134
Last seen: 11 years, 7 months
Re: Solve this problem, earn a cookie... [Re: Jii]
    #18406147 - 06/11/13 11:36 PM (11 years, 7 months ago)

I tried using both tap water and also purified water from a local dispensary.  Im not injecting water into anything... I soak the grain roughly 24 hours, heat to a simmer, let dry until its not sticking to my hands. Fill Jars/bags, PC, cool over night then inoculate...  my technique has worked for jars with grain to grain hundreds of times. i doubt my technique is the issue.  There's some sorta variable that im not seeing that's influencing my results.

Jii:
I've been more interested in mastering the sugar LC, but i did happen to knock up the bags straight with myc water, didn't take for me there either, but that same jar produced an LC that did product jars and eventually fruit.  I didn't play around with amounts though, and my guess would be that i used too much GLC, as i got a bacterial growth.  What would you usually use to inoculate say, 4 quarts?

and sorry for the slow response guys, i have limited access...

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: Solve this problem, earn a cookie... [Re: lgm0420]
    #18406176 - 06/11/13 11:46 PM (11 years, 7 months ago)

Hmmm.. well,
1) make sure what's growing in your cultures is mushroom mycelium in the first place.

2) sounds like (if I'm not mistaken) that nothing will grow on your grains right,(?)

Are you drying the grain surface prior to the PC run?  In a wet grain environment, mycelium will not grow and bacteria will get it. 


You could inoculate 4 quarts with a single drop of good culture.
Maybe your high volume of liquid is screwing your moisture level?

So I win the cookie?
We can share it in Guantanamo Bay together!


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OfflinePocketRevolution
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Registered: 12/22/09
Posts: 270
Last seen: 7 months, 26 days
Re: Solve this problem, earn a cookie... [Re: Oeric McKenna]
    #18406375 - 06/12/13 12:32 AM (11 years, 7 months ago)

How are you transferring the liquid from your liquid culture to the grains?  More specifically, what gauge needle is on your syringe?  If the syringe is too small, you may not be able to suck the mycelium into the syringe.  Also, do you have a piece of glass or some such thing inside the LC jar to break up the mycelium and agitate it into suspension before you draw out your sample?

Those are my best guesses at what could be amiss.

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Offlinelgm0420
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Registered: 02/18/09
Posts: 134
Last seen: 11 years, 7 months
Re: Solve this problem, earn a cookie... [Re: PocketRevolution]
    #18410298 - 06/12/13 07:32 PM (11 years, 7 months ago)

I guess there is no way to truly know if the mycelium in the liquid is good culture or not, since I cant get it to grow on grains... And im drying the grain to the point where it doesn't stick to my hand when I stick it in jars/bags.  Although I have played around with that in trying to get results... so I don't think thatll earn the cookie sadly.

As for the gauge of my needle, its what comes with a syringe from a vendor. I PC the syringes and re-use them.  I do my transfer in a glove box, in a still room, disinfected.  I check the syringe to see if I can visibly spot mycelium before I inject jars/bags.

The jar sits on a stir plate with a stir bar in the container, turns on about four or five times during the day and runs for 15 mins...

u see the frustration here? I cant find the hole...

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InvisibleOeric McKenna
LIFE CAPS


Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: Solve this problem, earn a cookie... [Re: lgm0420]
    #18411127 - 06/12/13 10:05 PM (11 years, 7 months ago)

If "anything" is visibly growing in your LC and you can't get "whatever it is" to grow on grain, you have a grain issue. Probably a moisture problem.  I've had growth issues in bags before too. Mostly commercially made ones I bought when I first started. They don't breathe too good.
Try jars dude.
  Rys seems to have more bacterial issues than other whole grains too. Though, if you're attentive to procedure, it'll go well.
  You could always try an eaiser grain, like wheat berries.


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OfflineRogerRabbitM
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Re: Solve this problem, earn a cookie... [Re: lgm0420]
    #18411194 - 06/12/13 10:18 PM (11 years, 7 months ago)

What you explain here is exactly why I won't use LC.  Not only are they slower than grains, you never know what you have until you use it.  With grains, you can take a whiff from each jar before you use it for spawn or grain to grain transfers.  This way, if you have a bad jar the problem stops right there.  With LC, you've usually spread the problem farther before it's discovered.
RR


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Offlinerustycobwebs
Green mold cultivator

Registered: 12/17/05
Posts: 799
Last seen: 9 years, 9 months
Re: Solve this problem, earn a cookie... [Re: RogerRabbit]
    #18412482 - 06/13/13 07:44 AM (11 years, 7 months ago)

Yea, LC is horrible. I like the idea, but in practice it is simply extra steps that require you to alter your preparation to accommodate that extra liquid.

I use a 14 gauge needle with LC, it's very different from trying to suck it up with an 18 gauge needle. 16 works pretty well, and that's I think what most vendors use.

Grain LC, or myc water, may be your best shot. Try sterilizing some water in a jar next time you run the PC? Then you'll have sterilized water you can pull out the injection port.

Is that how other people do the grain LC? Not to thread jack, but as far as liquid goes it seems like the safest route.


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Offlinelgm0420
Stranger
Registered: 02/18/09
Posts: 134
Last seen: 11 years, 7 months
Re: Solve this problem, earn a cookie... [Re: rustycobwebs]
    #18413880 - 06/13/13 02:20 PM (11 years, 7 months ago)

thanks for the input everyone...  I think my last attempt will be finding a different source of grain and using a different gauge needle.  Ive tried both bags and jars, so I don't think that's the issue.  As for sterilized water, that's how I normally do my grain LC's...

Really, the true issue here is my ego.  Instead of just cutting my loses im determined to make it work.  I guess one more go at it and then throw in the towel, time to make the smart decision.

Thanks guys

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