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 DZ
ranger
Registered: 01/25/09
Posts: 380
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 DZ's Inoculation Tek, WITHOUT GLOVEBOX! 
#10095229 - 04/02/09 02:36 PM (3 years, 1 month ago) |
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i really got tired of being confined by a glovebox so i started experimenting and came up with a method of inoculation that i enjoy much more and i've noticed no difference in contamination rates. this method is based on the assumption that you are inoculating from a syringe. i realize a glovebox is needed for some tasks like inoculating from spore prints, syringe making, agar, etc. but in my case this method has proved as effective as using a glovebox, so i'm going to stick with it, but you do what you feel is best, i just thought i'd share it as some may find it useful.
the key to this technique is motion efficiency!!!
lid preparation:
1) (for grain jar)instead of 2 medium sized holes, poke 3 into the lid.(2 for air exchange and 1 for the needle)
2) (for BRF jar)poke the standard 4 small holes, and add 2 medium sized holes for air exchange
* ^^^(not set in stone, we just need holes to breath and holes to inject)
3) then add a dab of silicone over every hole that is to be inoculated.(any kind of silicone should work, couple bux at lowes and such) allow to dry 12-24 hrs
*The silicone is considered self-healing so when we pull our needle from it we don't have to worry about un-sterile air going in.
*Lids are reusable so this step isn't necessary everytime
jar preparation:
1) the jar construction is: nutrients, a layer of tyvek(USPS = free tyvek envelopes), jar lid(up or down doesn't matter since tyvek will prevent from sealing), and then the screw-on ring
2) once jars are sterilized, allow to cool
now for inoculation:
*turn off air conditioner/heater we don't want it to come on and blow air around while we're working
*spray room with air sanitizer(oust or lysol) but no need to go nuts. i know it's tempting, but a few seconds has worked perfectly fine
1) remove foil used for sterilization
2) fold a paper towel in half and spray it with alcohol to where its thoroughly moist but not dripping.
3) place the moist alcohol paper towel on top of the jar and wipe it a bit, now let it sit on top of the lid (we're leaving it sitting on top of the jar to act as a contam barrier, we don't want anything landing on top of our silicone injection points before we inoculate)
4) now remove cap of your syringe and flame sterilize the needle (maybe within a foot or so from the jar you are getting ready to inject to), i like around 6 inches but NOT TOO CLOSE, ALCOHOL IS FLAMMABLE!!!
5) in 1 smooth motion, remove the needle from the flame, squirt a very small amount of solution to cool the needle(silicone doesn't like red-hot needles) and as quickly as possible slightly lift the paper towel and inject. this step should take a fraction of a second and should be as smooth as possible, the longer the needle is exposed to air the more chance of contamination we face. be careful needles are sharp and fingers are soft...
6) inject however much solution you would like
7) as you're pulling the needle out of the hole, wrap the paper towel around the needle, this assures the needle doesn't come in contact with an un-sterile environment so we won't need to flame sterilize between injection sites
8) (BRF tek)proceed onto the other 3 holes, always wiping injection site before injecting, and protecting needle from air contact
9) flame sterilize between jars, and continue until finished
i think i've eliminated as many contamination vectors as possible since the needle is only exposed to air for a very brief moment. this is the only method i use to inject from syringe anymore and i've found it to be very effective. i tried to be as thorough as possible so it probably sounds complicated but once you try it a few times you'll realize that it's not at all. enjoy.
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 NineInchNails
Stranger
Registered: 03/01/09
Posts: 547
Last seen: 1 month, 10 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: DZ]
#10100111 - 04/03/09 11:12 AM (3 years, 1 month ago) |
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Sure sounds reasonable to me.
Sure some may disagree, but I'm sure that MANY grow right out of their kitchen using less than sterile techniques with success. Just how successful they are ... I don't know. If it works, keep on doin it.
As long as you keep the paper towel saturated with fresh alcohol it should be fine.
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relaxing
kratomized
Registered: 04/01/09
Posts: 13
Loc: de south
Last seen: 6 months, 22 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: NineInchNails]
#10100262 - 04/03/09 11:53 AM (3 years, 1 month ago) |
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something is confusing to me, if the incubation holes are set with silicone for self healing injection sites, would not having extra ventilation holes negate that? -or- after you finish inoculating do you cover the entire jar top with a layer of aluminum foil?
I'm lucky to be in the light bulb business and plan to bath my inoculation area in germicidal light prior to inoculation. (if anyone researches this be sure to be clear on the hazards of germicidal light)
This tek sounds interesting as an addition, but I also like the inoculation tek called for simple minds or something?? I can't remember I saw a video on this site. I think that would be good if the area is bathed in germicidal prior to inoculation.
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DZ
ranger
Registered: 01/25/09
Posts: 380
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: relaxing]
#10100772 - 04/03/09 01:47 PM (3 years, 1 month ago) |
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well with the hole placement as long as the inoculation hole and the breather hole aren't too close(like within iono 1/4" ish) then it should be fine.
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bakamono
Wana match?
Registered: 10/27/08
Posts: 278
Last seen: 4 months, 24 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: DZ]
#10100818 - 04/03/09 01:58 PM (3 years, 1 month ago) |
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I knock up my jars w/o a gb just in my room, no prep besides sterilizing the needle and wiping my entry port with alocohol. 8 wbs jars this time around, 0 contams.
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DZ
ranger
Registered: 01/25/09
Posts: 380
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: bakamono]
#10100865 - 04/03/09 02:12 PM (3 years, 1 month ago) |
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Quote:
bakamono said:
I knock up my jars w/o a gb just in my room, no prep besides sterilizing the needle and wiping my entry port with alocohol. 8 wbs jars this time around, 0 contams.
are you doing spores or live inoculant ?
i'm so tempted to try my next G2G in open air next time since I'm probably using such a high ratio of healthy mycelium to grain that i think it won't make a difference as long as efficient motion is used.
anyone do G2G without GB or Flow Hood ?
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bakamono
Wana match?
Registered: 10/27/08
Posts: 278
Last seen: 4 months, 24 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: DZ]
#10100868 - 04/03/09 02:13 PM (3 years, 1 month ago) |
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spores, and lc's
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DZ
ranger
Registered: 01/25/09
Posts: 380
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: bakamono]
#10100897 - 04/03/09 02:19 PM (3 years, 1 month ago) |
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awesome, i figured the substrate would be more susceptible since the spores would take longer to take hold of the substrate. but i guess it doesn't really matter since most of the time (IME) if somethin funkys in there it usually pokes its head around fairly early.
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kiyote
Apprentice Grower
Registered: 07/08/08
Posts: 366
Loc: Near Chicago
Last seen: 1 month, 22 hours
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: DZ]
#10101160 - 04/03/09 03:07 PM (3 years, 1 month ago) |
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Nice writeup. Sterilization procedures depend on what your acceptable rate of failure/contams is. Personally, I use the exact same lid as you do, wipe it down with alcohol, wipe the needle down alcohol too, and then flame sterilize it, all in open air. I do repeat this with every injection but it does lead to an estimated 1 out of every 15-20 jars being contaminated. Not perfect, but since I really don't grow in that much bulk, it's acceptable...
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JPmagic
Welcome to the Dark Side
Registered: 03/02/09
Posts: 455
Loc: Midwest, US
Last seen: 1 year, 5 months
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: kiyote]
#10101219 - 04/03/09 03:19 PM (3 years, 1 month ago) |
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I honestly don't use a glove box when I do my inoculating. I just make sure that the only time the jar is exposed is for a quick 10 seconds. I've done 48 jars so far with 0 contams. Just make sure you don't leave lids or tinfoil off the top of the jar for very long and you should be fine. Yes, there is still a chance of contamination, but I have yet to see it, and will continue till I do.  PEACE!
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DZ
ranger
Registered: 01/25/09
Posts: 380
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: JPmagic]
#10101499 - 04/03/09 04:15 PM (3 years, 1 month ago) |
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thanks for your comments and for furthermore proving that a glovebox can be over-rated. i know it is considered next best thing to a flowhood but throughout beginner methods alternative methods to a glovebox seem to be left out. and being a beginner, i would've loved to know this info from the 'get-go'.
I should change my name to 'Glovebox Nazi'
(just wanted to make sure that none of this is misleading to anyone starting out, i only consider the glovebox unneeded when using syringes, other work certainly benefits from the use of a glovebox)
Thanks again guys and I appreciate your thoughts and experiences.
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samspade
Stranger
Registered: 06/08/09
Posts: 67
Last seen: 3 months, 6 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: JPmagic]
#10476678 - 06/09/09 10:36 AM (2 years, 11 months ago) |
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Instead of using a paper towel soaked in alcohol, just use alcohol wipes. Put a clean one over each jar and proceed.
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samspade
Stranger
Registered: 06/08/09
Posts: 67
Last seen: 3 months, 6 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: samspade]
#10476715 - 06/09/09 10:43 AM (2 years, 11 months ago) |
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Or one could use the "syringe-omatic" tek.
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Lennyk
D-O-L-E Dole
Registered: 04/22/08
Posts: 2,385
Loc: Near the Ground
Last seen: 10 months, 22 days
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Re: DZ's Inoculation Tek, WITHOUT GLOVEBOX! [Re: samspade]
#10476747 - 06/09/09 10:49 AM (2 years, 11 months ago) |
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Nice idea, but similar things have indeed been done before. Agar for one has had many modifications to the idea, one being the one in my signature with alcohol condoms.
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Stealth Lighting
Cubensis benefits beyond cluster headaches
Mush Extract! (You can even use Vinegar!)
Flame your needle in style with a sexy mini butane torch
~~~~~~~~~~~~~~~~~~~~~~~~~
What happens in the Romper Room, stays in the Romper Room.
~~~~~~~~~~~~~~~~~~~~~~~~~
All posts are written by the sex deprived helper monkey Curious George.
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