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OfflineCapless
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Registered: 06/25/06
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Bunk syringe, bad technique, or worse!?!
    #5854359 - 07/13/06 11:23 AM (17 years, 6 months ago)

Ok guys here is the story that is driving me to my wits end. I'll explain it from all angles possible.

Nine days ago I did a soak session for my WBS. I rinsed till the water ran clear, put it into the soaking water. Rinsed 24 hours later till the water ran clearn. I let it dry till no drippage could be seen and I put it in a holding pot to the side. I then snatched up 2 mycroporous bags filled each with two and a half(2.5) quarts a piece. I impulse sealed each one and then over the night & day I PC'd them one by one, each for an hour. Here is a possible error in my way, I'm only "pretty sure" my PC cooks at 15 PSI, I'm not completely sure on this hunch so I am acting on good faith here.

Any who I let each bag cool in the pot first, then I removed and let each cool in my clean room. My clean room is my bath room. I shut off the air flow to my bathroom, I consistantly sprayed the air with Lysol and Oust. I whiped down the counter with a nice alcohol solution and I washed my hands as well as I think I could. Once cool I brought one of my remaining B+ syringes from the refridgerator. I proceeded to shake it and *carefully* beat it against my hand to shake up the clumped spores inside. I continued on to steralize the syringe tip by heating the needle up to a red-hot state. After the needled cooled off into an alcohol bath it went untill I pulled it into the thick of a alcohol-drenched wad of napkin. I then injected roughly 5cc's of spore solution into each bag. I covered the injection site with a nice small round bandaid and I set my bags off into my closet. My room always maintains at least 75*F if not more.

All of that occured 7 days ago. Even today I have no growth.

Here are some reasons I'm somewhat skeptical about my *second* grow.

1) I'm not sure of my PC cooks at 15 PSI.
2) The B+ syringes came in from outside and the syringes themselves were very warm to the touch. I'm scared they might have died in the outside heat. This is the second grow to show no signs of mycellic growth.
3)I'm just being impatient.

My thoughts on the subject are that even if my PC didn't cook at 15PSI I think I should see at least some growth, even if it was to become contaminated. I'm really thinking the spores might have died due to the heat. The syringes were outside in a black mailbox on a 90*+ F day, and they were really warm when I put them into the refridgerator. If that is the case, I have made a live culture from a different set of spores that I ordered from the same company. They came in at a perfect temperature. I will most likely re-inoculate over the B+ if I have yet to see any growth by the time the LC is done.


--------------------
http://www.toolband.com/
"... This body holding me reminds me of my own mortality.
Embrace this moment. Remember, We are eternal.
all this pain is an illusion...."


Edited by Capless (07/13/06 11:33 AM)


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Invisibledecepticon
someone abovethe crowd

Registered: 07/10/06
Posts: 113
Loc: europa
Re: Bunk syringe, bad technique, or worse!?! [Re: Capless]
    #5854373 - 07/13/06 11:27 AM (17 years, 6 months ago)

your post got cut short man.
seems to happen a lot.

where those bags made to be impulse sealed before pc'ing?

and an hour each is probably not enough.


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OfflineCapless
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Re: Bunk syringe, bad technique, or worse!?! [Re: decepticon]
    #5854392 - 07/13/06 11:34 AM (17 years, 6 months ago)

Thank god I still had that saved on my cutting board so I could paste it.


--------------------
http://www.toolband.com/
"... This body holding me reminds me of my own mortality.
Embrace this moment. Remember, We are eternal.
all this pain is an illusion...."


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Invisibledecepticon
someone abovethe crowd

Registered: 07/10/06
Posts: 113
Loc: europa
Re: Bunk syringe, bad technique, or worse!?! [Re: Capless]
    #5854408 - 07/13/06 11:41 AM (17 years, 6 months ago)

that heat possibly could have hurt you.

no growth means no contams present as of this point.

so go start some other project and come back
in another week to look at the bags.

don't let the bags get too hot and wait.

if you see a problem, then begin to back track.
right now you're looking at dead spores or slow spores.
neither is that big of a deal.


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InvisibleTippinthru
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Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
Re: Bunk syringe, bad technique, or worse!?! [Re: Capless]
    #5854447 - 07/13/06 11:54 AM (17 years, 6 months ago)

Heat like you describe will often kill some spores, but not all.

Sounds like you PC'ed long enough & injected enough spore solution.

Inspecting, and handling bags daily, will impede growth.

I would have suggested making an LC, then injecting bags with it.

Advantage is: You will know if spores are viable (healthy LC).
You can use more inoculate & LC colonizes VERY quickly.

Just my take on your situation.



--------------------
Perfection is attained by slow degrees; it requires the hand of time...
[


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OfflineCapless
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Last seen: 14 years, 10 months
Re: Bunk syringe, bad technique, or worse!?! [Re: Capless]
    #5854457 - 07/13/06 12:00 PM (17 years, 6 months ago)

Thank you very much guys.


--------------------
http://www.toolband.com/
"... This body holding me reminds me of my own mortality.
Embrace this moment. Remember, We are eternal.
all this pain is an illusion...."


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Invisibleliamtheloser
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Registered: 06/07/06
Posts: 1,453
Re: Bunk syringe, bad technique, or worse!?! [Re: Capless]
    #5856028 - 07/13/06 08:56 PM (17 years, 6 months ago)

Also, no growth does _NOT_ mean no contams... If your pressure cooker doesn't reach 15psi, you most likely have a bacterial contamination that hasn't yet manifested in a visible manner.

If your PC does cook at 15psi, some of your spores MAY have been damaged in the heat, but the odds of the entire amount of spores dying is unlikely... After all, in nature, spores don't have air conditioned homes to live in while they're waiting to germinate.

But, since you did a multispore innoc instead of LC, wbs might take a while to show signs of colonization.

I hope everything turns out for you


--------------------


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OfflineCapless
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Re: Bunk syringe, bad technique, or worse!?! [Re: liamtheloser]
    #5856808 - 07/14/06 02:08 AM (17 years, 6 months ago)

Haha! Guys you will never believe how happy I am right now.

I'm sitting here in the early morning playing games. I thought to myself "Well I might as well re-inoculate those bags if they're just going to sit there with the LC's for the time being." I removed the first bag to my clean room, then I went to get the other. When I removed the second bag I was looking as hard as could for growth when I spotted a few grains on the bottom with that unforgettable white fuzzyness about them. I almost freaked out. I replaced that bag into its original position and to see the other. I checked the bottom on the other one and it is also showing mycelial growth!

Wish me luck for further on down the road!

Patience is golden in this hobby! Good things come to those who wait!


--------------------
http://www.toolband.com/
"... This body holding me reminds me of my own mortality.
Embrace this moment. Remember, We are eternal.
all this pain is an illusion...."


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