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Offlinelastround360
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Grain Spawn Question
    #27010870 - 10/29/20 05:41 PM (4 years, 2 months ago)

Hey guys. Making my first attempt at grain spawn currently and I believe I've made a vital mistake. I inoculated my spawn jars with LC about a week and a half ago. I was under the impression that rolling and shaking the grains around in the jar everyday to look for mycelium would not impact colonization. Now, a week and a half later, I see absolutely no mycelium in the jars and so I do some research and sure enough there is such thing as shaking too often. Does it sound like I need to start over with fresh grain or can I just re-inoculate with some more LC? I was literally shaking these jars everyday. Sometimes multiple times a day. I now understand I need to shake only right after inoculation and at around 30% growth. But what do you guys think?

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Offlinesonoramo
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Re: Grain Spawn Question [Re: lastround360]
    #27010904 - 10/29/20 06:00 PM (4 years, 2 months ago)

Funny you should ask that,...

I suppose it might be possible to kill off your inoculation by over-shaking it. If that's the case and you still have LC left, just re-inoculate. I did exactly that last weekend, when some of the jars I inoculated 4 days earlier didn't seem to "take."

In my case, I had used one 30ml syringe to inoculate 3 jars at a time (10ml each) in the first round, thinking to minimize the opportunity for contaminants getting into the LC. On the second round, I boosted to 15ml per jar, and made sure to agitate well so that each jar got equal amounts of mycelium.

Of course, you'll want to re-test your LC on agar, just to make sure it's still clean. :thumbup:

BTW: I just can't say enough about using long, blunt 16ga needles for grain inoculation. The long needles make it easy to flame sterilize the entire length of needle that goes into your jars, without risk of melting any plastic at the Luer lock.  :syringe:

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OfflinePBJ710
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Re: Grain Spawn Question [Re: sonoramo]
    #27011219 - 10/29/20 10:17 PM (4 years, 2 months ago)

Whats the source of the LC?  Normally it will at least try to start growing even with the grain getting beat to hell all the time - even if it can't expand, normally a few grains show colonization to some extent before the mycelium dies.

I wouldn't re-inoculate a jar that has known issues...thats just asking to bring problems to future grows.  Spend the $.07 on some more grain and write it off as a relatively cheap lesson learned.

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OfflineBrian Jones
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Re: Grain Spawn Question [Re: lastround360]
    #27011255 - 10/29/20 10:46 PM (4 years, 2 months ago)

Every time you shake a jar, it takes a couple days for colonization to recover.  Most people shake once or twice over the course of a grow.


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OfflineRoundabout
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Re: Grain Spawn Question [Re: Brian Jones]
    #27011269 - 10/29/20 10:57 PM (4 years, 2 months ago)

Stop shaking them and if nothing is happening after 3 or 4 days try innoculating again. I would be surprised if you could shake enough to prevent myc from taking hold entirely.

I often inspect my jars daily, but only do a vigorous shake once at maybe 30-50% colonized.

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Offlinesonoramo
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Re: Grain Spawn Question [Re: PBJ710]
    #27012074 - 10/30/20 12:51 PM (4 years, 2 months ago)

Quote:

PBJ710 said:
I wouldn't re-inoculate a jar that has known issues...thats just asking to bring problems to future grows.  Spend the $.07 on some more grain and write it off as a relatively cheap lesson learned.




I'm not sure if you posted in response to my comment. If contamination is the "known issue" then I'd have to agree :rolleyes: In my particular case, I had reason to think that the LC didn't get well-distributed in the syringe loads. That could explain how some jars really took off while others inoculated from the same syringe loads showed nothing after 3 days. This broth has a new Cube strain and I had forgotten to put a stir-bar in the LC jar when I processed it.

And BTW, all the jars are now showing fast growth and no sign of contamination. Also, no sign of contamination in the LC. :goodluck:

Another case where I'd agree "don't re-inoculate" is if you were to re-inoculate from a different LC. Growing is a continuous learning experience, and it will be hard to learn what strain does what if you mix strains in a jar.

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OfflinePBJ710
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Re: Grain Spawn Question [Re: sonoramo]
    #27013338 - 10/31/20 06:32 AM (4 years, 2 months ago)

That was addressed to the OP.  I wouldn't re-inoculate a jar (with the same LC or not) that didn't work the first time as you are not sure WHY it didn't work the first time.  You suspect it was the shaking, but there could be something else going on in it.  The reason I asked about the source of the LC is many people confuse LC with MS spore solution and that would dramatically effect your colonization consistency/times.  There could have been moisture or ??? issues that caused it to stall the first time as well.

Just trying to save you from having to re-do work.

Edited by PBJ710 (10/31/20 02:09 PM)

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