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 aenima
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Registered: 06/06/01
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Loc: UK
Last seen: 22 years, 9 months
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 Reduce contams with jars- extra step. 
#636024 - 05/19/02 03:50 AM (22 years, 11 months ago) |
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I haven't read anything about this techinique online or in any other teks.
I had been having an awful time with contamination in my jars, and was getting about 80+ contam rates. After reading about and experimenting (to a certain extent) i narrowed it down to where the contams were coming from.
After sterlising jars (using a pan on the stove as don't have a pressue cooker) i would take the jars out and let cool in a drawer in my bedroom. The jars it seemed were cooling down and sucking contam air into them, which was giving me the high contam rates. Leaving the jars in the pan on the stove to cool down wasn't an option, as live in a student house where not all people are tolerant to drugs.
Anyway i need something which would allow me to let the jars cool down anywhere without letting them get contamed, so i can up with the following extra steps in the preparation of jars.
Basically you make the jars as you normally do, i kinda follow MMGG with your substrate, then do the following.
Once you have the jars all prepared with your substrate, take a plastic sandwich bag ( i suspose you could use a oven bag, then you will be able to use a pressure cooker to sterlise as well) and place over the top of the jar make sure all the air is removeed from the bag and selotape in place. Don't want to place all the way over, just enough so you can selotape the bag to the sides of the jar.
Now sterilse as normal in a pan on the stove ( or with oven bag on top and in P.C, as a sandwich bag would melt in pressure cooker).
( As the jars sterilse they will release steam into the sandwich bag which will start to expand the sandwich bag. Once sterilsed and the jars are cooling they will suck the air from the sandwich bag back into the jars. This air will be clean and contam free. )
After an hour or so of sterlisation take the jars and let cool anywhere, the bags ontop will stop the jars getting contamed due to the reasons mentioned above.
now you can just inoculate as normal, if you want you can add an extra step to be even safer. I doubt this step is neccessary for most people, but i live in a student house which is dirty and need to be as safe as possible.
TO incoulate do the following.
Pull the sandwich bag up away from the jar, and inoculate as low down on the sandwich bag as you can, once inoculated psh the sandwich bag back down over the jars and selotape in place above the holes. Now there will be no way for contams to get in, as the only previous hole in the defnse would be the innoculation holes in the sandwich bag, but as they are selotaped above now, that hole is breached.
I am currently at 90 % colonized with 0% contam so far. Doing the jas the same as i always do, but with the added steps. This may be a coincedence, but i seriously think these steps have helped me out.
Will try and get some diagrams done to explain the above abit better later.
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HotDamn
newbie
Registered: 05/11/02
Posts: 42
Last seen: 22 years, 1 month
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Re: Reduce contams with jars- extra step. [Re: aenima]
#636026 - 05/19/02 03:54 AM (22 years, 11 months ago) |
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man...all this trouble. yeah, you could do that, or you could do it how i do, put electrical tape over the holes while sterilizing, and leave it on until you innoculate. then put more tape over the holes.
or you could spray your drawer with lysol...before and at the same time you put the jars inside.
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 Alien
Galactic Shaman
Registered: 11/14/99
Posts: 1,868
Loc: Jupiter's Moon
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Post deleted by Administrator [Re: aenima]
#636027 - 05/19/02 03:56 AM (22 years, 11 months ago) |
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-Alien 
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HotDamn
newbie
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Re: Reduce contams with jars- extra step. [Re: HotDamn]
#636028 - 05/19/02 03:56 AM (22 years, 11 months ago) |
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oh and for innoculation, use an oven as a flow hood...you do have one of those dont ya?
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Hippie3
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Registered: 11/06/99
Posts: 3,090
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Re: Reduce contams with jars- extra step. [Re: HotDamn]
#636039 - 05/19/02 04:12 AM (22 years, 11 months ago) |
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hey hot damn, take a burning stick of incense and put it in your oven and see where the smoke goes--
an oven is no substitute for a flowhood or even a decent glovebox.
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Alien
Galactic Shaman
Registered: 11/14/99
Posts: 1,868
Loc: Jupiter's Moon
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Post deleted by Administrator [Re: Hippie3]
#636047 - 05/19/02 04:19 AM (22 years, 11 months ago) |
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-Alien
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HotDamn
newbie
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Re: Reduce contams with jars- extra step. [Re: Hippie3]
#636050 - 05/19/02 04:21 AM (22 years, 11 months ago) |
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wha? i think your example is misleading..the oven tek has worked quite well for me, and i actually thought it was you that invented the thing. I only open the door a large crack and wear gloves to prevent burning my hand from the rising heat, and innoculate.
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ChromeCrow
one ancient mutha
Registered: 02/21/02
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Re: Reduce contams with jars- extra step. [Re: Alien]  1
#636106 - 05/19/02 05:46 AM (22 years, 11 months ago) |
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a homemade glovebox is doing me right. a tupperware box with a plexi glass peice epoxied on the lid, holes in the side, with the sleeves of an old rain coat epoxied all around, and rubber dishwashing gloves epoxied to the end of the sleeve. its so airtight, that i have to hurry on flame steralizing, or i use up the air, and kill the flame. ive been doing agar work in it, and have got 0% contams with it
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ISO: Orissa, Malabar, z strain
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indicaz
Lysol God
Registered: 04/12/02
Posts: 580
Loc: My Chair
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Re: Reduce contams with jars- extra step. [Re: ChromeCrow]
#636168 - 05/19/02 07:04 AM (22 years, 11 months ago) |
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regarding a glovebox, if your poor and cant afford a big ole plastic box and gloves
Ive found that as long as most of your work area is isolated from direct air traffic your pretty good, Ive used a carboard box lysoled down, basically ive found that as long as I isolate my general work area, wipe it down with a bleack solution and spray generously with lysol and work inside any type of box I never get contams..
I do own a glovebox but I never use the sucker, I have 3 dogs and am a cigarette smoker and my house is far from sterile! But ill tell you what my work area always gets wiped down heavily before use, during use and after. I spray before, During and after. Like I said though if ya cant afford a glovebox a Carboard Box is even good, by isolating your work area you will be more likely not to lose cakes to contams...... Im no expert buit this has worked for me.
Oh P/S The Oven Tek Dont Use It, It has been proven that when innoculating air is actually sucked back into the jars, I beleive theres pics of the proof on here somewhere....
Laterz
Indicaz
Later
Indicaz
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phrozendata
Carpal Tunnel
Registered: 04/23/00
Posts: 5,015
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Re: Reduce contams with jars- extra step. [Re: Hippie3]
#636186 - 05/19/02 07:21 AM (22 years, 11 months ago) |
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I think Hippie3 is refering to something Anno posted a while ago...and just to proove that the search engine does work (you n00bs just need to learn how to use it) I found it for you all:
He also said that some of the smoke goes all the way to the bottom of the jar! Other experiements concluded that the oven tek, in fact, is not good at all. It's funny...i've been using it for a wihle now and just switched to open air innoculations (in my room sprayed with lysol which is kept partially sterile but I still live in it ya'know?) and have had no problems. I'll probably be too much of a bitch to try making a syringe in open air...I guess i'll have to construct a glovebox or something *shrugs* 
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"There's only one corner of the universe you can be certain of improving and that's your own self. So you have to begin there, not
outside, not on other people" - Aldous Huxley
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cbull
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Posts: 363
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Re: Reduce contams with jars- extra step. [Re: aenima]
#636271 - 05/19/02 08:37 AM (22 years, 11 months ago) |
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dude when jus befor u put the cap on the jars to sterlize use tinfoil make it tight and put it inbetween the jars and the lid when your ready to inject wipe off the tin foil with what ever. if u use metal jars lids and poke a hole in 4 spots, u can wipe the tin foil through the holes. under the lid where u poke the hole jus make sure there no metal edges so when u screw the lid on over the tinfoil befor sterlizing the tin foil wont rip
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FAPA - Member Since -5-21-2002
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HotDamn
newbie
Registered: 05/11/02
Posts: 42
Last seen: 22 years, 1 month
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Re: Reduce contams with jars- extra step. [Re: cbull]
#636679 - 05/19/02 03:23 PM (22 years, 11 months ago) |
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Is that an empty jar? how high was the heat? was innoculation performed by holding the jar in hand over a not fully opened oven door?
Edited by HotDamn (05/19/02 03:25 PM)
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aenima
member
Registered: 06/06/01
Posts: 194
Loc: UK
Last seen: 22 years, 9 months
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Re: Reduce contams with jars- extra step. [Re: HotDamn]
#636962 - 05/19/02 08:14 PM (22 years, 11 months ago) |
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have tried oven tek and get high contam rates, plus can't always get to the kitchen due to aforementioned house mate who isn't exactly pro drugs and it isn't woth my time arguing with them.
It isn't that much extra work, maybe 45 secs altogther, i am more than happy to put in 45 secs of extra work for 0% contamination so far.
Tried having tin foil over the jars, but that still results in contams for me, as i guess it isn't a complete seal.
Bleach sprayed my drawer, but that didn't stop the contams.
The method mentioned above, does. Just thought i'd post it so others could perhaps use it who are in a similar position to me.
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I've been wallowing in my own confused, insecure dellusions
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DERRAYLD
Constructus
Registered: 05/13/02
Posts: 12,982
Loc: South Africa
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Re: Reduce contams with jars- extra step. [Re: aenima]
#637012 - 05/19/02 09:14 PM (22 years, 11 months ago) |
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I lock myself in the bathroom, own a house so I can take as long as I want.
Spray the entire room down with 50/50 bleach and water.
The room sure as hell sterilises. No chance it can smell that clean and not be 
Let everything settle and open sterilised bottles.
I use consol jars, self sealing lids.
When I take them out the pot they are vacuum sealed.
I`ve left them for 3 days and not a single sign or contamin.
All jars are inoculated and still no contam.
been about 5 days now.
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Northernsoul
Your Reality
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Posts: 2,290
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Re: Reduce contams with jars- extra step. [Re: Alien]
#637037 - 05/19/02 10:28 PM (22 years, 11 months ago) |
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For the cool down part: I just leave them inside the pressure cooker...as for the oven thing, i dont bother...i just clean my bathroom real real good, spray it down witha a whole can of lysol and put a hepa filter in there for an hour...when im in there i just make sure there arent any drafts what so ever . put a towel at the foot of the door etc...100% contam free so far. You dont even need the hepa filter, ive done it without the filter b4 and that worked 100% too...its just an added precautionary toy.
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When it comes
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Northernsoul
Your Reality
Registered: 11/17/01
Posts: 2,290
Loc: Inner Eye
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Re: Reduce contams with jars- extra step. [Re: Northernsoul]
#637038 - 05/19/02 10:31 PM (22 years, 11 months ago) |
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PS: oh ya, a dust mask and always flaming my needle too and wipe hands with alcohol constantly
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When it comes
I'll know, I know
Just take my clothes and leave
And I'll be gone
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DERRAYLD
Constructus
Registered: 05/13/02
Posts: 12,982
Loc: South Africa
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Re: Reduce contams with jars- extra step. [Re: Northernsoul]
#637046 - 05/19/02 10:52 PM (22 years, 11 months ago) |
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I just keep my 50/50 bleach solution handy and just spray the old hands a couple times and viola.
Or I just keep the sink full of 50/50 bleach.
I use a gas mask and hair net of sorts. That`s only to keep the dreads bleach free otherwise they don`t like the bleach.
I stink of bleach for the day but that`s cool
as long a no contam.
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Hippie3
mycotopiate
Registered: 11/06/99
Posts: 3,090
Loc: mycotopia.net
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Re: Reduce contams with jars- extra step. [Re: DERRAYLD]
#637128 - 05/20/02 01:49 AM (22 years, 11 months ago) |
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good lord, 50/50 bleach water is way too strong, just overkill.
hard on yer lungs, too.
just a teaspoon or 2 per gallon is enough to sanitize surfaces.
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DERRAYLD
Constructus
Registered: 05/13/02
Posts: 12,982
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Re: Reduce contams with jars- extra step. [Re: Hippie3]
#637134 - 05/20/02 02:03 AM (22 years, 11 months ago) |
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it`s not the lungs I`m concerned about, my eyes have been burning for the past 2 days lol oooops
Nah it`s all cool.
But I can tell it worked, my damn bathroom is probably still sterilised from last night lol
Ppl have been saying as well that you shouldn`t sterilise jars with rubbers down and tightened bcause it doesn`t actually sterilise.
I can tell you now I left my jars for 3 almost four days and I had not a single spot or contam????????????????
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aenima
member
Registered: 06/06/01
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Loc: UK
Last seen: 22 years, 9 months
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Re: Reduce contams with jars- extra step. [Re: DERRAYLD]
#644266 - 05/25/02 02:38 AM (22 years, 10 months ago) |
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100% contam free, 95% colonized in two weeks from syringe.
All of the above methods for 100% contam free sound like more than an extra 45 seconds of work to me.
*to post below* yes that is an option, but to read the first post, that was not an option, hence this idea.
*said once again* this works for the conditions i am in, all the other suggestions have been tried and failed, hence why this was thought up, and alot of the time the suggestions given here do not meet the conditions i am in.
*wonders to self* do people actually read threads before posting
*answers self* obviously not.
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I've been wallowing in my own confused, insecure dellusions
Edited by aenima (05/25/02 04:48 AM)
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