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OfflineMajickMuffin
Edible Cult


Registered: 05/28/14
Posts: 4,345
Loc: North
Last seen: 2 years, 8 months
Is my 'knock up' method Sterile? Pf tek
    #22264152 - 09/20/15 10:36 AM (8 years, 4 months ago)

So yesterday I had 20 Whole wheat Flour jars, followed a tek to the T, was quite simple.
Pot-sterilized, followed a tek to the T, was quite simple.

I believe I prepared them 100% correctly so lets not talk about that, lets assume they are good.

I knocked up 10 with 1 MS syringe of Cubensis, the other 10 with 1 MS of Pan Cyans(so excited).

(The reason im posting this is for experienced people to rate my success rate based on my inoculation method, please and thank you :smugjerry:)

My method:
I did this in open air, but the air was still, no windows no drafts (maybe a slight one that I could not feel).
I put the syringe tip onto the MS syringe in a drawer quickly but smoothly.
Used a Butane torch to flame sterilize the tip (which only took like 1 second till it was RED). Allowed it to cool for around 30s, then wiped the tip with an Alcohol swab.
A few seconds to allow the Alcohol to evaporate then injected 1 jar, 2 holes. I injected into 2 holes without doing anything to the Syringe.
After 1 jar, I would wipe the Syringe tip with an Alcohol swab, gently set down, put micropore tape over the holes on the knocked up jar (yes it has a verm layer, im just being safe!)
Then I would start on the next jar.

SO, in between jars, I would wipe the syringe tip with an alcohol swab. I did not flame sterilize in between each jar...it seemed unnecessary, time consuming, and possibly negative or problems could happen. I had 20 jars and 2 syringes...

Ive knocked jars up using much less sterile methods than that and have had about an 80% success rate.

Thanks for any input, suggestions, :thisisheavyman::pedrophile::havesomescience::cheersyoufuck:


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Offlineshroombasa
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Registered: 05/02/15
Posts: 364
Loc: 'Merka
Last seen: 3 years, 3 months
Re: Is my 'knock up' method Sterile? Pf tek [Re: MajickMuffin]
    #22264201 - 09/20/15 10:49 AM (8 years, 4 months ago)

Multispore is always a crap shoot regardless.  Open air inoculation with syringes really isn't a big deal, IMO.  I think the biggest issue is wiping down with alcohol instead of flame sterilizing.  Rubbing with alcohol after flame sterilization is one of those topics where people get really uncivil.  Personally, I'd just flame sterilize and forget the alcohol rub.


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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City Flag
Re: Is my 'knock up' method Sterile? Pf tek [Re: shroombasa]
    #22264320 - 09/20/15 11:19 AM (8 years, 4 months ago)

why not 70% or 90% EtOH/IspOH -- it worked fine for me.

i mean, that's what doctors and nurses do before giving a shot..

i never bothered with flame sterilisation but i don't see why it would hurt

i *did* work in back of the bunsen burner, trick i picked up working in a lab

all in all, it sounds good to me


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(Avatar is Maxxy, a character by Mizzyam, RIP)


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Offlinesaintburger
Helper


Registered: 09/18/14
Posts: 469
Last seen: 1 month, 18 days
Re: Is my 'knock up' method Sterile? Pf tek [Re: micro]
    #22264383 - 09/20/15 11:33 AM (8 years, 4 months ago)

Because sanitizing is not the same as sterilizing. Why would you sanitize a sterile needle? It is only hampering your sterile technique if anything...


--------------------
Set goals for yourself, or wander around aimlessly.

"One day Alice came to a fork in the road
and saw a Cheshsire cat in a tree.
'Which road do I take?' she asked.
'Where do you want to go?' was his response
'I don't know', Alice answered.
'Then', said the cat, 'it doesn't matter.'"


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Offlineshroombasa
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Re: Is my 'knock up' method Sterile? Pf tek [Re: micro]
    #22264385 - 09/20/15 11:33 AM (8 years, 4 months ago)

Sorry, that's one thing I forgot to add.  If it works for you and you're satisfied with your contam rate, go with it. 

When I started, I would do the exact same thing.  Flame sterilize first, then wipe after each jar.  My contam rate wasn't that bad at all.  It's only after I did several G2G transfers and realized that I had a contam I didn't spot that I switched to all flame sterilization.  I went from like a 10% contam rate to close to 0%.  The best justification I've heard for flame sterilization over alcohol is that alcohol won't clean the inside of the syringe, whereas red hot metal will.


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OfflineMajickMuffin
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Registered: 05/28/14
Posts: 4,345
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Last seen: 2 years, 8 months
Re: Is my 'knock up' method Sterile? Pf tek [Re: shroombasa]
    #22264425 - 09/20/15 11:43 AM (8 years, 4 months ago)

After the first flame sterilization the inside of the syringe should remain sterile no matter how many jars you go through, because liquid is shooting out of it keeping the contents of the syringe tip from the sterile contents inside the syringe.

The only thing I was worried about was the outside of the syringe tip because every time I would get through a jar I would sometimes touch the syringe tip onto the jar lid, although it should be sterile, but also from open air.

You guys answered my questions, thanks a lot all!


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Offlinesaintburger
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Registered: 09/18/14
Posts: 469
Last seen: 1 month, 18 days
Re: Is my 'knock up' method Sterile? Pf tek [Re: MajickMuffin]
    #22264473 - 09/20/15 11:55 AM (8 years, 4 months ago)

You should flame sterilize between each jar in PF tek to avoid cross contamination.
After flame sterilization the needle shouldn't touch anything, it should go straight to the jar.

Open air is no big deal but you might as well get around to building an SAB. If you're serious about growing and want to learn more things, you'll need one in no time.

Flame sterilization is always more effective than wiping a needle with alcohol. Fire sterilizes. Alcohol sanitizes. Sterile is better than sanitized.


--------------------
Set goals for yourself, or wander around aimlessly.

"One day Alice came to a fork in the road
and saw a Cheshsire cat in a tree.
'Which road do I take?' she asked.
'Where do you want to go?' was his response
'I don't know', Alice answered.
'Then', said the cat, 'it doesn't matter.'"


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Offlineblackout
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Registered: 07/16/00
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Re: Is my 'knock up' method Sterile? Pf tek [Re: micro]
    #22264599 - 09/20/15 12:23 PM (8 years, 4 months ago)

Quote:

micro said:
why not 70% or 90% EtOH/IspOH -- it worked fine for me.

i mean, that's what doctors and nurses do before giving a shot.



Doctors wipe your skin, not the needle. Wiping the skin is like wiping a SHIP.


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OfflineKizzle
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Re: Is my 'knock up' method Sterile? Pf tek [Re: shroombasa]
    #22264661 - 09/20/15 12:34 PM (8 years, 4 months ago)

Quote:

i mean, that's what doctors and nurses do before giving a shot..



They wipe the injection site, not the needle. Alcohol wipes become contaminated very quickly so unless you're using a fresh presterilized wipe each time you'd be better off not wiping it all.


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Offlinevagblast
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Registered: 10/04/10
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Re: Is my 'knock up' method Sterile? Pf tek [Re: Kizzle]
    #22264882 - 09/20/15 01:15 PM (8 years, 4 months ago)

A sab, Flame sterilization between each jar, squirt a tiny bit out to cool needle no alcohol wipe and u will have success every time.


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OfflinecronicrFacebook
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Re: Is my 'knock up' method Sterile? Pf tek [Re: vagblast]
    #22264914 - 09/20/15 01:23 PM (8 years, 4 months ago)

Quote:

vagblast said:
A sab, Flame sterilization between each jar, squirt a tiny bit out to cool needle no alcohol wipe and u will have success every time.



:whathesaid:


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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City Flag
Re: Is my 'knock up' method Sterile? Pf tek [Re: saintburger]
    #22265361 - 09/20/15 02:52 PM (8 years, 4 months ago)

Quote:

saintburger said:
Because sanitizing is not the same as sterilizing. Why would you sanitize a sterile needle? It is only hampering your sterile technique if anything...




Makes sense.

We always cleaned the injetion site...


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)


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