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OfflineMoose420
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Made a mistake and learned from it. Don't use dirty needles
    #28630826 - 01/22/24 09:20 AM (5 days, 22 hours ago)

So I'm just getting started in this awesome hobby and made a mistake someone else might learn from.

So about 2 weeks ago I got some spores and inoculated a few bags of rice. Which are doing great (I think)
About 4 days ago I also tried the PF tek method with a few jars. I did 3 jars of 1 strain and 3 of another. 1 strain seemed to take off after only 2 days and I thought that was odd from what I read considering I started from spores. But what I thought was mycelium is definitely something else as it's turning green.

I did not sterilize the needle between jars for that atrain. The other was a new needle. I initially did but not for the others. I think what happened was there must of been a piece or rice or something in the needle that started to rot in-between inoculation dates. These jars are getting tossed.




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OfflineSkropi
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420]
    #28630888 - 01/22/24 10:35 AM (5 days, 21 hours ago)

Not sterilising the needle between inoculations is a mistake, but did you sterilise the jars before inoculation? I ask because if you follow the PF Tek to the letter, I believe you should have more than 97% success.


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OfflineMoose420
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Skropi]
    #28630909 - 01/22/24 10:54 AM (5 days, 20 hours ago)

Yes jars were sterilized in a pressure cooker.


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OfflineHelloImBob
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420] * 1
    #28631041 - 01/22/24 12:49 PM (5 days, 18 hours ago)

Quote:

Moose420 said:
So I'm just getting started in this awesome hobby and made a mistake someone else might learn from.

So about 2 weeks ago I got some spores and inoculated a few bags of rice. Which are doing great (I think)
About 4 days ago I also tried the PF tek method with a few jars. I did 3 jars of 1 strain and 3 of another. 1 strain seemed to take off after only 2 days and I thought that was odd from what I read considering I started from spores. But what I thought was mycelium is definitely something else as it's turning green.

I did not sterilize the needle between jars for that atrain. The other was a new needle. I initially did but not for the others. I think what happened was there must of been a piece or rice or something in the needle that started to rot in-between inoculation dates. These jars are getting tossed.








Trichoderma is a common issue and yes grows faster and starts out white before turning green.

Flame sterilizing needles is common practice but seeing as you have 3 jars with major trichoderma growth are you sure it's not the spore syringe that is dirty?

Is it all the same strain that grew trichoderma?


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"You appear to be talking about boosting tryptamine content in mycelium by amending LC with....whatever your amendments are. I have to say I'm a tad disappointed that you're addressing us with the shorthand of a 13 yo girl who's texting her besty for make-up tips, instead of proper English, which causes me to have doubts."


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Invisibleelasticaltiger
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: HelloImBob] * 1
    #28631050 - 01/22/24 01:00 PM (5 days, 18 hours ago)

It's interesting that the green all seems to come from the inoculation points.

How long did you sterilize and what are you using for gas exchange?

Your summation that it's the needle COULD be correct but the fact that it seems to be growing from where the solution dropped, to me, means that it was actually the solution itself or incomplete sterilization which caused the areas that were hydrated by the solution to germinate first.


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OfflineHelloImBob
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: elasticaltiger]
    #28631074 - 01/22/24 01:32 PM (5 days, 18 hours ago)

Yeah that's what I was wondering if it's the spore solution because the rest of the jar looks fine.

Seems like a lot more trichoderma than the needle itself would carry as well.

if the needle was dirty, I could see one jar for sure, let's say you lost all the trichoderma spores flushing the needle and contacting the material possibly cleaning it on your sterilized medium but 3 jars with major trichoderma?

Looks like the spore solution to me.


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Quote from Stipe-n-Cap

"You appear to be talking about boosting tryptamine content in mycelium by amending LC with....whatever your amendments are. I have to say I'm a tad disappointed that you're addressing us with the shorthand of a 13 yo girl who's texting her besty for make-up tips, instead of proper English, which causes me to have doubts."


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OfflineMoose420
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420]
    #28631094 - 01/22/24 02:06 PM (5 days, 17 hours ago)

Yes all the same strain. But the 2nd time using that syringe.


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OfflineMoose420
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: elasticaltiger]
    #28631105 - 01/22/24 02:13 PM (5 days, 17 hours ago)

Hi
I sterilized for 45 minutes in the pressure cooker after it came up to pressure. I'm using a layer of vermiculite as well as micropore tape over the holes.
The other 3 jars I did of a diffrent strain/syringe seem to look good. There is nothing growing yet.

Here are a few pictures of the bags I did about a week earlier. Don't see anything but what I think is mycelium in those. These are from the spore solution in question.

And a few pics of the other jars/strain








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OfflineHelloImBob
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420] * 1
    #28631181 - 01/22/24 03:17 PM (5 days, 16 hours ago)

I get that you reused the needle, but it's sounding like one of your sryinges has mushroom spores and trichoderma spores mixed together.

If it's all the same strain then it's sounding like your inoculant is dirty.


--------------------
Quote from Stipe-n-Cap

"You appear to be talking about boosting tryptamine content in mycelium by amending LC with....whatever your amendments are. I have to say I'm a tad disappointed that you're addressing us with the shorthand of a 13 yo girl who's texting her besty for make-up tips, instead of proper English, which causes me to have doubts."


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OfflineLogicaL ChaosM
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420] * 1
    #28631195 - 01/22/24 03:30 PM (5 days, 16 hours ago)

Yeah its really important to clean that needle tip inbetween jar inoculations. It can pick up trich spores from the air inside a SAB during that time.


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OfflineHelloImBob
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: LogicaL Chaos]
    #28631217 - 01/22/24 03:53 PM (5 days, 15 hours ago)

Sorry my bad, I need to learn to read.

Didn't see the part that said these are from the spore solution in question.

I'm an idiot


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Quote from Stipe-n-Cap

"You appear to be talking about boosting tryptamine content in mycelium by amending LC with....whatever your amendments are. I have to say I'm a tad disappointed that you're addressing us with the shorthand of a 13 yo girl who's texting her besty for make-up tips, instead of proper English, which causes me to have doubts."


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OfflineMrJong
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: HelloImBob] * 1
    #28631241 - 01/22/24 04:33 PM (5 days, 15 hours ago)

Clean, flame, poke, next jar, clean, flame, poke, (...) and if you're not done with the syringe and plan on reusing it, push just a drop out of it on a tissue or something, flame and close the cap.

You can flame inbetween injections on the same jar but it quickly becomes redundant if your technique is proper.

I try to keep the cap sterile by only opening the pack it comes in and the syringe it's on afterwards in my SAB.


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OfflineMoose420
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: MrJong]
    #28631702 - 01/23/24 04:52 AM (5 days, 2 hours ago)

Thanks for all the replies. Loving this hobby


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OnlineMwj12977
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Re: Made a mistake and learned from it. Don't use dirty needles [Re: Moose420] * 1
    #28631709 - 01/23/24 05:08 AM (5 days, 2 hours ago)

I’ve forgotten to sterilize here and there when I’m doing a bunch at a time and still end up with clean jars. Typically around 95% are good.  I think your issue was a bad syringe to begin with.


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