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Offlinerisky80
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my first inoc...
    #22067254 - 08/09/15 01:36 PM (8 years, 5 months ago)

I used a glove box.  Flame sterilized out of the box then when in the box i would use alcohol soaked paper towel to clean and then inject.  I did 10 jars.  I feel i was very clean throught. I over injected kinda hard to see theough the plastic.  Has this happened to you before. Basically am hoping for the best i never did this before.  I hear people do open air and have great success.


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Offlineinked4life
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Re: my first inoc... [Re: risky80]
    #22067365 - 08/09/15 01:59 PM (8 years, 5 months ago)

the main issue is air movement, open air innoc is ok, sterize the work space, close windows, elimate drafts of air and use same precidure for innoc jars with flaming needle n wipe off with alcohol wipe. ive never had an issue


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InvisibleInocuole
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Re: my first inoc... [Re: risky80]
    #22067369 - 08/09/15 01:59 PM (8 years, 5 months ago)

Quote:

risky80 said:
I used a glove box.  Flame sterilized out of the box then when in the box i would use alcohol soaked paper towel to clean and then inject.  I did 10 jars.  I feel i was very clean throught. I over injected kinda hard to see theough the plastic.  Has this happened to you before. Basically am hoping for the best i never did this before.  I hear people do open air and have great success.




So you sterilized, and then you wiped dirty paper towels all over it before you inoculated? Never use a sanitizer after flame sterilizing, you're just getting it dirtier.  Mold spores can survive alcohol but not flame.  That's bad.  Also if you have gloves attached to your box that's even worse.

Might work out but if I knew I just made that many mistakes I would start over.  I don't even know what you mean by "over injected" but I'm certain that's the least of your problems.


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Invisiblemustangbob3
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Re: my first inoc... [Re: Inocuole]
    #22067401 - 08/09/15 02:07 PM (8 years, 5 months ago)

i have only ever nocc'd up in open air with syringes:shrug:

SAB i only use for g2g innocs

i do it open air but be quick and flame between each jar/bag.

never had a problem in 10 years.:shrug:

i would more worry about whats in the syringe that your squirting in the sterile bag.

dirty spores or risky LC.

so really if your going inject you cant be sure of cleaness anyway so as i see it a flame here and there and wipe down the bag and be quick is as good as it gets or needs to be for me.

it dont matter how good your procedure if the spores/lc is funky


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OfflineFreeWorldOrder
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Re: my first inoc... [Re: mustangbob3]
    #22067695 - 08/09/15 03:25 PM (8 years, 5 months ago)

Quote:

mustangbob3 said:
i have only ever nocc'd up in open air with syringes:shrug:

SAB i only use for g2g innocs

i do it open air but be quick and flame between each jar/bag.

never had a problem in 10 years.:shrug:

i would more worry about whats in the syringe that your squirting in the sterile bag.

dirty spores or risky LC.

so really if your going inject you cant be sure of cleaness anyway so as i see it a flame here and there and wipe down the bag and be quick is as good as it gets or needs to be for me.

it dont matter how good your procedure if the spores/lc is funky




I use a SAB for all inoculations, agar work, G2G etc. The whole concept of a SAB is to have space to work in where the air is still and not swirling around with unwanted mold spores & bacteria. There are very good resons a SAB is used and recommended....


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InvisibleMad Season
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Re: my first inoc... [Re: inked4life]
    #22067745 - 08/09/15 03:32 PM (8 years, 5 months ago)

Quote:

inked4life said:
the main issue is air movement, open air innoc is ok, sterize the work space, close windows, elimate drafts of air and use same precidure for innoc jars with flaming needle n wipe off with alcohol wipe. ive never had an issue



but it's not a still environment.. do all that and tell me the room wouldn't be covered in dust after 6 months. That's how much the air is moving. Also wipe after?! How the hell have you not had a problem?!?! Let's see you in a few years :lol:

@OP why don't you just straight up make an SAB and stop using alcohol. Flame only.


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Edited by Mad Season (08/09/15 03:35 PM)


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OfflinetaGyo
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Re: my first inoc... [Re: Mad Season]
    #22067816 - 08/09/15 03:44 PM (8 years, 5 months ago)

Quote:

taGyo said:
Alochol SANITIZES, flames STERILIZE. Sterilization means NO germs, mold can survive sanitation.
Don't wipe your needle with alcohol after flame sterilization:

Quote:

swatt_haze said:
wiping a needle with alcohol after flame sterilizing is like rubbing shit on your hands after you washed them and eating a burger.





Flame until the needle is red hot. It will turn black first (because you're using a lighter), then slowly turn red.

Flame sterilize between each JAR, not each inoculation hole. .25 cc a hole for 1cc total each BRF jar.
Grains is 1CC as well.

After flame sterilization gently push down on the plunger. It'll shoot out steam until a drop of liquid comes out. Let the drop gently fall and then push your needle through the hole, making sure to angle it TOWARDS THE GLASS. I look down and check to make sure my needle is against the glass sometimes but you can feel it.

If you push and it feels difficult and nothing's coming out gently pull your needle upward while shooting. BE CAREFUL, this will immediately loosen up your needle and if you're pushing too hard you'll shoot way too many CCs into a hole.




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Invisiblemustangbob3
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Re: my first inoc... [Re: FreeWorldOrder]
    #22067852 - 08/09/15 03:50 PM (8 years, 5 months ago)

Quote:

FreeWorldOrder said:
Quote:

mustangbob3 said:
i have only ever nocc'd up in open air with syringes:shrug:

SAB i only use for g2g innocs

i do it open air but be quick and flame between each jar/bag.

never had a problem in 10 years.:shrug:

i would more worry about whats in the syringe that your squirting in the sterile bag.

dirty spores or risky LC.

so really if your going inject you cant be sure of cleaness anyway so as i see it a flame here and there and wipe down the bag and be quick is as good as it gets or needs to be for me.

it dont matter how good your procedure if the spores/lc is funky




I use a SAB for all inoculations, agar work, G2G etc. The whole concept of a SAB is to have space to work in where the air is still and not swirling around with unwanted mold spores & bacteria. There are very good resons a SAB is used and recommended....




yeah i know mate

me personally i dont get the point if using MS Ioccs
the risk/chance of infection from the contents of the syringe is much much much greater then the risk from a needle being in open air for literally a second.

so if using MS I dont bother. im lazy

but yes i do agree 100% and everyones homes are different ect
i just have never had a problem doing as when i way the risk up the extra effort aint worth it.

and a syringe can still pick up bacteria from the air in a SAB anyway its just less likely with good technique.(even the still air has not been cleaned and is full of bacteria ect that stays in suspension)

for anyone else and most so with people who are starting out do it all in the SAB, 1 less thing to go wrong:thumbup:
and then at least you will be able to narrow down the cause of contams if it should arrise:thumbup:

but i do think the risk of doing it in open air is very small in comparison to the chance the MS syringe itself is clean.


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OfflinetaGyo
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Re: my first inoc... [Re: mustangbob3] * 1
    #22067860 - 08/09/15 03:52 PM (8 years, 5 months ago)

You can innoc in open-air.

When you flame the needle you don't just kill everything that lands on it but a phenemonon called thermophoresis begins, which repels all microscopic bacteria with heat from the needle.

Relax, different folks different strokes. As long as we all get clean spawn.

:thumbup:


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InvisibleInocuole
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Re: my first inoc... [Re: taGyo]
    #22067878 - 08/09/15 03:54 PM (8 years, 5 months ago)

That stops working the second you eject a drop to cool the needle which you're supposed to do more than a few inches away from your inoculation point. :shrug:

Why ever inoc in open air when we know better?  I know you tried that open air agar shit.


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OfflinetaGyo
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Re: my first inoc... [Re: Inocuole] * 1
    #22067883 - 08/09/15 03:55 PM (8 years, 5 months ago)

Quote:

Inocuole said:
That stops working the second you eject a drop to cool the needle which you're supposed to do more than a few inches away from your inoculation point. :shrug:

Why ever inoc in open air when we know better?  I know you tried that open air agar shit.



I don't knock shit unless it's in my SAB.

I was imagining he was putting the needle in hot and cooling it in the substrate, which works just fine as well.


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Invisiblemustangbob3
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Re: my first inoc... [Re: Inocuole]
    #22067919 - 08/09/15 03:59 PM (8 years, 5 months ago)

agar and stuff is a diff story.

i do it as like i said i weigh the risks.

and the fact is the MS syringe is the biggest risk factor.

so if using them i dont go the whole hog!

and if you do it in a SAB still how do you stop the airbourne shit just like when in open air? imo a SAB also requires good technique for it to preform at its best. how you move your hands, do transfers, and g2g while disturbing the air as little as possible.

no amount of spraying ect will remove them lots hang in the humidity of the air and will never drop out. and bleach spray wont get em all niether will oust.

so less risky but still risk of the same thing is there.
the weak link imo is the MS syringe and most contams if they happen will be from this and not air contact with the needle providing your grains sterile.


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



Edited by mustangbob3 (08/09/15 04:05 PM)


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OfflinetaGyo
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Re: my first inoc... [Re: mustangbob3]
    #22067971 - 08/09/15 04:12 PM (8 years, 5 months ago)

Quote:

mustangbob3 said:
and if you do it in a SAB still how do you stop the airbourne shit just like when in open air?




By having the air in my still air box be still after a nice iso bath?


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InvisibleJacobStorm
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Re: my first inoc... [Re: taGyo]
    #22068033 - 08/09/15 04:23 PM (8 years, 5 months ago)

I've knocked a lot of jars in open air, flame sterilizing the needle then alcohol wipe the needle...
Thought that's what we're supposed to do. I haven't had a sing jar contaminate yet.


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Invisiblemustangbob3
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Re: my first inoc... [Re: taGyo]
    #22068036 - 08/09/15 04:24 PM (8 years, 5 months ago)

even still air will be full of bacteria and unless you are going to hepa filter the air you aint getting rid of them!

the risk is greatly reduced but the air will still be teaming with bacteria even in a still air box that you will be moving the needle around in.

and beside my main point was the risk from whats in a ms syringe greatly out weighs the risk of injecting in open air.

but it is 1 more risk so i do agree but i still dont bother lol :thumbup:


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InvisibleInocuole
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Re: my first inoc... [Re: mustangbob3]
    #22068047 - 08/09/15 04:26 PM (8 years, 5 months ago)

Quote:

mustangbob3 said:
even still air will be full of bacteria and unless you are going to hepa filter the air you aint getting rid of them!

the risk is greatly reduced but the air will still be teaming with bacteria even in a still air box




If you're familiar with the principles of a still air box, the moisture(iso or water) against the floor and walls of the SAB draws airborne contaminants into it over the course of the 30 minutes you're supposed to let it sit with all your air turned off in the house.  By the time it's run its course there very well should be a pocket of sterile air in there, the rest still being significantly cleaner than open air.


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OfflinetaGyo
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Re: my first inoc... [Re: mustangbob3]
    #22068054 - 08/09/15 04:28 PM (8 years, 5 months ago)

Quote:

taGyo said:
Alochol SANITIZES, flames STERILIZE. Sterilization means NO germs, mold can survive sanitation.
Don't wipe your needle with alcohol after flame sterilization:

Quote:

swatt_haze said:
wiping a needle with alcohol after flame sterilizing is like rubbing shit on your hands after you washed them and eating a burger.





Flame until the needle is red hot. It will turn black first (because you're using a lighter), then slowly turn red.

Flame sterilize between each JAR, not each inoculation hole. .25 cc a hole for 1cc total each BRF jar.
Grains is 1CC as well.

After flame sterilization gently push down on the plunger. It'll shoot out steam until a drop of liquid comes out. Let the drop gently fall and then push your needle through the hole, making sure to angle it TOWARDS THE GLASS. I look down and check to make sure my needle is against the glass sometimes but you can feel it.

If you push and it feels difficult and nothing's coming out gently pull your needle upward while shooting. BE CAREFUL, this will immediately loosen up your needle and if you're pushing too hard you'll shoot way too many CCs into a hole.




Ok. So.

Flame kills all the germs instantly. The way iso works is by penetrating the cell walls of things and then evaporating, killing the germs. This is a process that takes about two minutes. If you flame sterilize, kill everything, and then add shit onto it you're actually doing the opposite of what the flame intended.

If you haven't seen a contam, great. IMO you haven't been growing long enough if you haven't seen a contam. I'd switch up my procedure before it comes to bite you in the ass.
Quote:

mustangbob3 said:
even still air will be full of bacteria and unless you are going to hepa filter the air you aint getting rid of them!

the risk is greatly reduced but the air will still be teaming with bacteria even in a still air box



Honestly any time you knock up a jar you risk contams :shrug:

The goal of the SAB and other non-open-air environments is to greatly REDUCE that risk. That's not to say that just because I use an SAB I will never get aerial contams floating into my jars but if open air is 1 in 25 chance then SAB is 1 in 100.

I'd take that 1 in 100 any day :shrug:. I'm even underestimating the SAB I believe. Some people rarely see contams using them and have been growing for years.

Also, what Inoc said. You will have contams in there, sure, but it's not even near the same amount that are floating around in open-air.


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InvisibleMad Season
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Re: my first inoc... [Re: Inocuole]
    #22068082 - 08/09/15 04:34 PM (8 years, 5 months ago)

Also particles still have a weight to them. They're still effected by gravity. Why would it just float in the air? Everything settles but air currents won't let it. In a STILL air box, you should have no air currents. The moisture everywhere catches everything and the air will settle. This is why I can get 99% success with open agar dishes. This is what happened when I opened a dish FOR LITERALLY 1 SECOND in a bathroom behind the shower curtain with the air off for months:



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Invisiblemustangbob3
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Re: my first inoc... [Re: Inocuole]
    #22068096 - 08/09/15 04:35 PM (8 years, 5 months ago)

thats a pipe dream

you cant really believe that surely.:smile: even hepa filtering air dosent remove all contaminates!! and thats the best you can get!

some/lots will stay in suspension i assure you.

even using a flow hood has risks and requires good technique to be really efficient.

its laughable you think the air in a SAB is clean. leave it as long as you want but it will never be so.


Quote:


Honestly any time you knock up a jar you risk contams :shrug:

The goal of the SAB and other non-open-air environments is to greatly REDUCE that risk. That's not to say that just because I use an SAB I will never get aerial contams floating into my jars but if open air is 1 in 25 chance then SAB is 1 in 100.




yes thats why i said i agree 100% as it REDUCES chances but explained why i dont bother when using MS.
:thumbup:

please dont take what im saying out of context:)

g2g ect im right there with you:)

but the odd time i use unsafe MS (which i have good luck with anyway) i just dont do it, i agree i prob should and i agree with what you are saying:thumbup:

its just a small risk after the huge risk i already made when using MS in the first place.

so i expect that it might not pan out and dont rely on it.:smile:
like i said if your new to the hobby and want the most you can out of it remove all risk that you can and use a SAB for everything :thumbup:
and that way if anything goes wrong you can easily narrow the cause down to the cause.





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Edited by mustangbob3 (08/09/15 04:46 PM)


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InvisibleMad Season
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Re: my first inoc... [Re: mustangbob3]
    #22068131 - 08/09/15 04:40 PM (8 years, 5 months ago)

With a good technique and a killer eye, it doesn't reduce it. It gets rid of contaminations. I've never had a contam with proper sterile technique. It's always been me fuckin up hard core.


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