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InvisibleRobMarley420
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Registered: 05/01/05
Posts: 12,554
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How do I make sure my syringe 100% sterile?
    #4422054 - 07/19/05 02:33 AM (18 years, 7 months ago)

I need to re-inocculate 5 jars and I need to make sure that my needle and syringe is 100% sterile.
    I inoculated 3 golden teacher jars and 5 B+ jars. I had some  Golden Teacher spore solution left over in a syringe from my last grow. It was enough to knoc' up 3 jars. I sterilized the needle with an alchohol flame for about 15 secs, then wiped it off with hydrogen peroxide. All 3 of those jars got contamanated! :mad: :frown:  I knoc'ed up 5 jars with B+ from a brand new syringe, the package was never opened. Never touched the needle or attempted to sterilize. All 5 of those jars show no signs of contams! :smile: But they are going slow as fuck compared to GT. :frown:  So I'm 100% sure that the Golden Teachers were contamed becuase of the reused syringe, not because of my sterilization proccess (pressure cooking and inoculation environment).
    All of my B+ jars are going SLOW. 3 show tiny signs of growth but only at 1 injections site, the other 2 show nothing at all. I'm going to re-inject all of these jars to get some mycelium growing at all 4 injection sites on all 4 jars. I need to know how to sterilze my needle to make sure it won't contam my precious cakes. I sterilized my GT syringe with an alcohol flame and then wiped it down with h2o2. There was still a little bit of air in the syringe that was pushed out before i inoculated. Is this where the contams came from? Any help would be great!

These or my B+
jars



These pics were taken 8 days after inoculation.

Here on 2 of the 3 contamed GT jars


I ordered my spores from spores101.com. My GT syringes have about 10 times more sprores than my B+. The GT syringes have huge clumps of spores but one of my B+ syringes has a very small clump and the other has barely any at all! Is this the reason why the Golden Teachers were taking off and the B+ are so slow? And also on my last grow all of my Golden TEacher jars grew from all 4 injection sites, on this grow my B+ are only growing form 1 site (if any) Is this cause the syringe didn't have much spores in it? Why did they hook it up so SHADY on the B+ syringes?


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Edited by RobMarley420 (07/19/05 02:36 AM)

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OfflineBenKenobi
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Registered: 05/22/05
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Re: How do I make sure my syringe 100% sterile? [Re: RobMarley420]
    #4422338 - 07/19/05 03:55 AM (18 years, 7 months ago)

Ive reused syringes b4 no contam as yet. I just give the syringe an initial squeeze to get what is in the needle out then i flame it for 5sec with a BIC lighter and give it a good rub with a soaked alcohol towel to wipe the black residue off. Then i wipe down my jars with alcohol and insert needle, i redo this process for each jar. I also do all my work inside of a glove box.

I cant see how a resued syringe can be your problem if you are sterilising the needle prior to use. Unless of course u sucked in some air after originally using them but i doubt u would of done that. For them to be the reason of contamination then a contam needs to get inside the syringe, providing you are using very sterile procedures with everything else. Might just of been a case of bad luck.

I personally wouldnt worry bout re-injecting more spores if u are worried about possibly getin a contam, as long as the jars arnt dryin out the myc will soon enough end up getin around the jar just might take a couple weeks.

GL anyway.


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Old Ben

I beleive that my job in mappin the impulse pathways of a single neuron can allow us to open a mecca into the quailitative functions of the human geno...

Please support your local free spore ring by sendin back prints from successful grows, FSRE

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OfflineKalix
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Registered: 03/20/05
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Re: How do I make sure my syringe 100% sterile? [Re: BenKenobi]
    #4422375 - 07/19/05 04:33 AM (18 years, 7 months ago)

I wouldn't put more spores in the B+, you are inviting bacterial contams that way, by throwing off the water content...


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My Unitarian Jihad Name is: The Shotgun of Sweet Reason

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InvisibleA3eyedfish
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Registered: 05/06/99
Posts: 1,454
Loc: Wut some people Call Par...
Re: How do I make sure my syringe 100% sterile? [Re: BenKenobi]
    #4422380 - 07/19/05 04:37 AM (18 years, 7 months ago)

pour yourself about 4 ozs of h202, remove the needle from the partially used syringe, dip the syringe end into the peroxide, shake it off, seal the syringe end with a hot glue gun. When you are ready to to use it again, sterlize the needle & syringe, twist off the hot glue & replace the needle, you should be ready to go


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InvisibleFooMan
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Registered: 02/02/05
Posts: 8,957
Loc: Earth Flag
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Re: How do I make sure my syringe 100% sterile? [Re: BenKenobi]
    #4422410 - 07/19/05 05:07 AM (18 years, 7 months ago)

Quote:

BenKenobi said:
i flame it for 5sec with a BIC lighter and give it a good rub with a soaked alcohol towel to wipe the black residue off. Then i wipe down my jars with alcohol and insert needle, i redo this process for each jar. I also do all my work inside of a glove box.






flame+alcohol+glove box=  :burnbabyburn:

Be careful! Lysol too.


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Quick WBS Prep

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OfflineBenKenobi
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Registered: 05/22/05
Posts: 342
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Last seen: 10 years, 10 months
Re: How do I make sure my syringe 100% sterile? [Re: FooMan]
    #4422421 - 07/19/05 05:21 AM (18 years, 7 months ago)

lol yer i flinch every time i spark it dont worry, hasnt happened yet thou so fingerscrossed.


--------------------
Old Ben

I beleive that my job in mappin the impulse pathways of a single neuron can allow us to open a mecca into the quailitative functions of the human geno...

Please support your local free spore ring by sendin back prints from successful grows, FSRE

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Offlinelemon_lw
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Registered: 10/17/04
Posts: 3,622
Loc: That Way
Last seen: 16 years, 6 months
Re: How do I make sure my syringe 100% sterile? [Re: BenKenobi]
    #4422517 - 07/19/05 07:22 AM (18 years, 7 months ago)

it lopks like you have tape on your jars. if you do take it off and there will be no need to re innoc unless you messed up the water content.


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In the belly of the Leviathan, one can either despair and perish, or be cheerful and persevere.-Dean Koontz

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