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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,870
Last seen: 2 months, 7 days
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Re: Where are my contams coming from? [Re: Ghatti]
#20585320 - 09/18/14 07:30 PM (10 years, 3 months ago) |
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Maybe you should give it one last attempt and try a grow bag. They can be completely colonized and even produce the first flush before being exposed to anything outside the sterile environment of the bag. It'd eliminate any possibility of contamination being introduced during the spawn run because they don't need to be spawned.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Kizzle] 1
#20585374 - 09/18/14 07:43 PM (10 years, 3 months ago) |
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I've been inoculating jars of sterilized grain/substrate mixes with liquid inoculant in my SAB and fruiting them out of the jars. They stay in sterile conditions until its time to fruit.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585407 - 09/18/14 07:52 PM (10 years, 3 months ago) |
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A grow bag would be equivalent to a grain jar with a SHIP I would think. Problem is I don't have a clean syringe and as far as your suggestion Muda, I wouldn't trust an LC if I can't successfully inoculate a grain jar with an agar wedge with any consistency.
I'm going to let the jars go that I knocked up once I made the hepa filtered positive pressure box. It probably won't work but it only cost me 8 bucks. Thought was to hopefully knock back some of the funk I was bringing in when I put my arms in. Looking back at it it has a very very low chance of changing anything.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Ghatti] 1
#20585426 - 09/18/14 07:56 PM (10 years, 3 months ago) |
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I don't use LC I make non nutritive Liquid inoculant with Agar wedges and sterilised water. Second link in my sig.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585454 - 09/18/14 08:06 PM (10 years, 3 months ago) |
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Sadly I don't own a compatible blender. Even then I wouldn't trust opening a jar to pour it in or anything else really at this point.
I really should just get a syringe and do ms grows but none of my jars have ships so I'd still have to crack lids anyways so I'd likely be in the same boat as using wedges.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20585474 - 09/18/14 08:09 PM (10 years, 3 months ago) |
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Pf tek
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Ghatti]
#20585492 - 09/18/14 08:13 PM (10 years, 3 months ago) |
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I open my jars for every inoculation, Even with syringes. I don't like the Idea of my needle coming into contact with a SHIP which is only sanitised and not sterile. It's a small risk I know but it still doesn't sit right with me.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585504 - 09/18/14 08:14 PM (10 years, 3 months ago) |
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-------------------- The Basics
A little civility goes a long way
The Noob Forum
The Hammock Hangers' Forum
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585515 - 09/18/14 08:16 PM (10 years, 3 months ago) |
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I don't like the idea either, hence why none of my jars have them. But I've about resigned myself to the fact that contams are getting in during inoculation be it off of my gloves, my agar, or just floating around.
So cracking lids is likely a no-go unless I'm downstream on a hood.
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
Posts: 8,438
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Re: Where are my contams coming from? [Re: Kizzle]
#20585649 - 09/18/14 08:42 PM (10 years, 3 months ago) |
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Quote:
Kizzle said: Maybe you should give it one last attempt and try a grow bag. They can be completely colonized and even produce the first flush before being exposed to anything outside the sterile environment of the bag. It'd eliminate any possibility of contamination being introduced during the spawn run because they don't need to be spawned.
... was gonna say. There are places that do the work for you.
Their rates are reasonable.
Fuck PM me and I'll prep some media for you to inoculate. You have to promise me you'll only use it for oysters though.
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber
The Shmuvbox. - The Old TC's Like it
Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger! No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit.
"The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT
Pastywhite's Easy Agar Tek (PastyPlates)
Tiger Drop Video Demos By munchauzen
Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson
EZEKIEL 23:20
Edited by elasticaltiger (09/18/14 08:42 PM)
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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That's all I attempt to growz. I don't think its the media. I really think its happening when I inoculate so even if you sent me something sterile, it wouldn't be once I got a hold of it lol.
I don't think its the culture cause when I spawned the jars individually half were good. If it was the culture I woulda thought everything would go bad.
I don't think its the filters or media since I just switched to sfd's and I pc'd a group of jars and let them sit for 2 weeks with no contams.
So that really just leaves stuff getting in when I inoculate right?
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,870
Last seen: 2 months, 7 days
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585716 - 09/18/14 08:54 PM (10 years, 3 months ago) |
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Quote:
MudaFuka said: I open my jars for every inoculation, Even with syringes. I don't like the Idea of my needle coming into contact with a SHIP which is only sanitised and not sterile. It's a small risk I know but it still doesn't sit right with me.
Put a piece of tape over the SHIP before sterilization When you want to inoculate the jar flame sterilize your needle. While still red-hot lightly touch the end of the needle to the center of the tape killing any contaminants in that spot on contact, hold it there for a few seconds as the needle cools so you don't damage your port and press it through.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Kizzle]
#20585748 - 09/18/14 09:02 PM (10 years, 3 months ago) |
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I don't use syringes very often anymore but that's a good tip.
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,870
Last seen: 2 months, 7 days
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Re: Where are my contams coming from? [Re: MudaFuka]
#20585794 - 09/18/14 09:11 PM (10 years, 3 months ago) |
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Yeah I don't either. I suppose because of the hatred I've acquired of contaminated LCs and spore syringes but not because it doesn't work.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: Kizzle]
#20585953 - 09/18/14 09:38 PM (10 years, 3 months ago) |
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So the control plates I opened for 10 secs, got bacteria in one of em. Guess I'll wait a few days and see if any mold germinates.
Hell bacteria I could deal with lol
If there's bacteria there's prolly spores on there too. I have the sterile air blowing down so I probably wafted my hand too close over the dish and the eddy's blow crap in.
I wonder since my air purifier uses a blower (centrifugal fan) could I not just put the 2 brand new hepas side by side and build a flange to fit it.
It wouldnt be laminar but it should be sterile and if I turned my tub on its side with the open end facing me and cut the bottom out and mounted the filters in I wonder if it wouldn't give me a nice little space to manipulate it seeing as it has walls and a roof?
Edited by Ghatti (09/18/14 09:50 PM)
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Mdahmer
Aloysius devadander abercrombie



Registered: 04/05/14
Posts: 2,516
Last seen: 7 years, 5 months
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Re: Where are my contams coming from? [Re: Ghatti]
#20586080 - 09/18/14 09:59 PM (10 years, 3 months ago) |
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i think you just need to maybe look at your SAB and make sure theres nothing wrong or odd about it and just work at your sterile tech. if you spawned all those jars by themselves and only half failed im guessing your just not very smooth with innoc jars and whatnot. i practiced with my old agar plates in my sab while watching TV doing transfers over and over then going to jars over and over until i felt like a machine. smoking a lot of herb helps.
i wouldnt quit growing after dragging this thread along, its just not right
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: Mdahmer]
#20586113 - 09/18/14 10:05 PM (10 years, 3 months ago) |
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Lmao you have a point. Well I have a spare tub so I may try my idea anyways, what can it hurt at this point in the game anyways?
I'd love to work on my sterile technique but I honestly don't see how I can improve it. I feel like I've had so many failures that everything is just smothered in mold spores and even if I wash up I still get nasty by the time I go in my sab.
I do the same process as I do with my agar plates and they turn out fine 99% of the time.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Mdahmer]
#20586115 - 09/18/14 10:05 PM (10 years, 3 months ago) |
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It took me a lot of practice to get good at sterile work. I fucked up well over 100 plates before I got 1 without bacteria.
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Mdahmer
Aloysius devadander abercrombie



Registered: 04/05/14
Posts: 2,516
Last seen: 7 years, 5 months
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Re: Where are my contams coming from? [Re: MudaFuka]
#20586132 - 09/18/14 10:09 PM (10 years, 3 months ago) |
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yea me too with the bacteria, it was terrible. ive only seen mold twice that i remember except on old ass unwrapped plates but i would be for sure seeing bacteria, it was a matter of having clean enough growth away from it to get a good sized chunk for my jar lol.
i got a bunch of bacteria growing right now from a few different clones actually..fucking shit
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: MudaFuka]
#20586140 - 09/18/14 10:10 PM (10 years, 3 months ago) |
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My plates are usually clean, my jars however look clean but grow green once spawned to cvg and introduced to fae.
Hey that sounded spiffy.
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