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OfflineDinkinFlicka84
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Determining if I have bad spawn...huge issues I've been perplexed with.
    #21275435 - 02/14/15 11:58 PM (9 years, 3 months ago)

So I have 10 out of 65 jars that never colonized. Maybe I missed them when I was inoculating.
I also had 3 jars go green during colonization about 2 weeks in (60% colonized).
My thought is, if these jars with no myc in them never got mold, but 3 jars out of 65 that DID, does that mean I could possibly have bad inoculate? Or does everyone experience a jar or 2 going bad during colonization from a GLC?

My technique is the same every time in plunge a syringe into an injection port.
I PC The syringe, use 70% alcohol on everything from my gloves to the injection ports.
I wear a mask and do this in my bathroom with the A/C off as to minimize any moving air.

Also, I just started dunking my spawn right before I mix with bulk. I was told to dunk for 10-15 min, but originally I soak my WBS for 12-24 hours, so why wouldn't I need to soak/dunk the spawn for a similar amount of time to rehydrate the grains again?


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OfflineForgottenPassword
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: DinkinFlicka84]
    #21275443 - 02/15/15 12:04 AM (9 years, 3 months ago)

sounds like youre doing pretty good to me. its not like youre in a lab. of course youll get a few contam. the other jars may still colonize, were missed, or just really unlucky it takes 2 spores together to germinate,

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OfflineKizzle
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: ForgottenPassword]
    #21275664 - 02/15/15 02:02 AM (9 years, 3 months ago)

You should be flame sterilizing the needle as well before each inoculation.


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Invisiblegrainbrain
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: Kizzle]
    #21275762 - 02/15/15 04:06 AM (9 years, 3 months ago)

Quote:

Kizzle said:
You should be flame sterilizing the needle as well before each inoculation.




This, also did you test your GLC prior to knocking up 65 jars?


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Invisibleblindingleaf
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: grainbrain]
    #21275773 - 02/15/15 04:15 AM (9 years, 3 months ago)

u could have a bad innoculant, and the jars that appear healthy just aren't showing mold yet.

or the ones that did get green may have had a lapse in filter, OR when u injected, u brought something in that took a bit of time to germinate and go green.

when people dunk grains before spawning, they are re-hydrating the mycelium, so a short soak is fine.  when u soak grains for grain prep, ur hydrating the grain with its tough outer hull, so a longer soak is called for.

like kizzle said, FLAME the needle.  its really important.  heat is the only thing that destroys the organisms we are looking to destroy during culture work.  whether u use a $1.00 shitty crack lighter, or a $300 bacticinerator, you need to get the needle red hot before injecting or aspirating.


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OfflineDinkinFlicka84
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: blindingleaf]
    #21279720 - 02/15/15 08:19 PM (9 years, 3 months ago)

Is there a way I can put some of my inoculate from a LGC onto a petri dish to see if it's bad? I'm about to get into agar, but I wanted to see if there is something I can do right now to determine if I have bad spawn before I inoculate a bunch of jars.


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InvisiblebodhisattaMDiscordReddit
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: DinkinFlicka84]
    #21279731 - 02/15/15 08:24 PM (9 years, 3 months ago)

Quote:

DinkinFlicka84 said:
Is there a way I can put some of my inoculate from a LGC onto a petri dish to see if it's bad? I'm about to get into agar, but I wanted to see if there is something I can do right now to determine if I have bad spawn before I inoculate a bunch of jars.



yes

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OfflineDinkinFlicka84
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: bodhisatta]
    #21280669 - 02/15/15 11:26 PM (9 years, 3 months ago)

Good to know. Is there a specific way I do this? I'm never done agar before, but I'm trying to do this without getting into agar right away. I want to order some petri dishes and whatever else I need to simply see if I have bad spawn. If i can tell whether or not my spawn is good, then I won't need to get into agar. I have been doing GLC for a while and it works 85% of the time.


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InvisibleBig Bear
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: DinkinFlicka84]
    #21280709 - 02/15/15 11:34 PM (9 years, 3 months ago)

Put a drop or two on a petri and see how it looks after a few days at room temp :thumbup:


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OfflineDinkinFlicka84
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: Big Bear]
    #21280878 - 02/16/15 12:24 AM (9 years, 3 months ago)

Just so I don't screw this up...
I need to use my still air box with the prep that goes with it, then add a few drops to a petri dish from my LGC. Then wait a few days to see if it grows mold?

I should've done this a long time ago...
I've always been curious about my spawn being bad because I would have semi-constant problems with contamination. So, I could make a LGC, then squirt a few drops on a petri dish to check if it's good, then I would know in a couple days that the culture is ok to use to inoculate multiple jars. Yes?


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OfflineKizzle
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: DinkinFlicka84]
    #21280986 - 02/16/15 01:26 AM (9 years, 3 months ago)

You'll want to make a streak with it on the agar using a sterile cotton swab or inoculation loop.


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InvisiblebodhisattaMDiscordReddit
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Re: Determining if I have bad spawn...huge issues I've been perplexed with. [Re: Kizzle]
    #21283531 - 02/16/15 03:30 PM (9 years, 3 months ago)

Quote:

Kizzle said:
You'll want to make a streak with it on the agar using a sterile cotton swab or inoculation loop.



or a spread plate with a glass rod with a 90ยบ bend.

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