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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: PsiloBeast]
#20382248 - 08/06/14 11:12 AM (9 years, 7 months ago) |
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I'm not leaning towards it because I've used different syringes, many different plates that I was sure were clean.
I always blamed a dirty syringe or a super stealthy contam on every single plate I had which is unlikely
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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: Ghatti]
#20382264 - 08/06/14 11:14 AM (9 years, 7 months ago) |
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Well I have to pin it down to one thing now since I have reviewed hardcore my sterile procedure, my filters, my jars and lids, my cultures, etc. The only 2 things I have left that I can think of is my g2g and replacing poly with sfd's for good measure though I doubt its the issue.
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PsiloBeast
Stonesun Wannabe
Registered: 05/23/14
Posts: 2,517
Loc: The Wild
Last seen: 9 years, 6 months
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Re: Where are my contams coming from? [Re: Ghatti]
#20382294 - 08/06/14 11:22 AM (9 years, 7 months ago) |
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Lol QFT by dos ounce though
...contams are rarely from one single source. Contams are all around us by the trillions in numbers, all the time. To assume there is a single reason why your jar are failing may be a failure in and of itself so to speak. I get about a 5-10% contam rate so 1/20 jars I make or so (from MS syringe) contams. Don't know why. Don't know where. Don't really care all I do is try to be a little bit cleaner next time. Re read Teks and procedures, and try again.
You might be thinking wayy too hard about this whole situation. Insecurity breeds negativity.
-------------------- Whosoever is delighted in solitude is either a wild animal or a god... -Aristotle A COMPLETE GROW LOG (ATL# 7) BEAST STYLE LINKS Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...
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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: PsiloBeast]
#20382346 - 08/06/14 11:34 AM (9 years, 7 months ago) |
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I would agree with you and be ecstatic if I had a 5-10% contam rate right now but I don't. It's close to 100% when I combine quarts and make a tub. But much lower when I do single quart trays, likely the good trays are from the receiving jars that were poured first or second before I replaced the lid many times. Which goes to tell me its not random or just unlucky, there is a fundamental flaw somewhere and this is one thing that hasn't changed since I've been troubleshooting and tweaking/changing other things and I still have the exact problem.
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PsiloBeast
Stonesun Wannabe
Registered: 05/23/14
Posts: 2,517
Loc: The Wild
Last seen: 9 years, 6 months
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Re: Where are my contams coming from? [Re: Ghatti]
#20382414 - 08/06/14 11:50 AM (9 years, 7 months ago) |
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For sure man, I really hope you find what it is!!
-------------------- Whosoever is delighted in solitude is either a wild animal or a god... -Aristotle A COMPLETE GROW LOG (ATL# 7) BEAST STYLE LINKS Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...
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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: PsiloBeast]
#20382457 - 08/06/14 12:00 PM (9 years, 7 months ago) |
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Yeah if changing my culture AGAIN, switching to sfd's, and doing my g2g differently don't help then my fungi friendship will have ended and I'll leave yall to it as I'm out of things I can change and try.
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Kizzle
Misanthrope
Registered: 08/30/11
Posts: 9,866
Last seen: 21 days, 4 hours
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Re: Where are my contams coming from? [Re: Ghatti]
#20382565 - 08/06/14 12:23 PM (9 years, 7 months ago) |
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Didn't you rule out G2Gs already? That's always the first thing I suspect.
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PsiloBeast
Stonesun Wannabe
Registered: 05/23/14
Posts: 2,517
Loc: The Wild
Last seen: 9 years, 6 months
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Re: Where are my contams coming from? [Re: Ghatti]
#20382702 - 08/06/14 01:01 PM (9 years, 7 months ago) |
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Quote:
Ghatti said: Yeah if changing my culture AGAIN, switching to sfd's, and doing my g2g differently don't help then my fungi friendship will have ended and I'll leave yall to it as I'm out of things I can change and try.
That's not the greatest attitude to have in mycology my friend!
-------------------- Whosoever is delighted in solitude is either a wild animal or a god... -Aristotle A COMPLETE GROW LOG (ATL# 7) BEAST STYLE LINKS Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...
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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: PsiloBeast]
#20382858 - 08/06/14 01:40 PM (9 years, 7 months ago) |
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I am not one to quit easily but I've been at this for many months with no end in sight and have tried everything I can think of.
And kizzle I thought I had ruled out g2g by spawning nothing but masters but a look back told me I was mistaken. So I'm revisiting the issue wholeheartedly.
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Dos Ounce
Fighter of the Nightman
Registered: 09/07/12
Posts: 873
Loc:
Last seen: 3 years, 8 months
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Re: Where are my contams coming from? [Re: Ghatti]
#20383193 - 08/06/14 03:22 PM (9 years, 7 months ago) |
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don't take this the wrong way
BUT
I feel like if you're following proper sterile procedures then you wouldn't be having any problems.
maybe you're doing something small that you don't even realize is the problem, so you haven't mentioned it. Not saying that's the case, but if it were you wouldn't know otherwise.
you should revert back to pf tek cakes and work your way back up to bulk after spawning cakes to trays successfully, that kind of thing.
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TheEaglesGift
The Nagual
Registered: 04/10/11
Posts: 10,554
Loc: Ixtlan, Mexico
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Re: Where are my contams coming from? [Re: Dos Ounce]
#20383270 - 08/06/14 03:41 PM (9 years, 7 months ago) |
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Quote:
maybe you're doing something small that you don't even realize is the problem, so you haven't mentioned it.
This.
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blojo02184
Big Red
Registered: 05/15/13
Posts: 3,527
Loc: Maine
Last seen: 1 month, 17 days
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Re: Where are my contams coming from? [Re: Dos Ounce]
#20383463 - 08/06/14 04:17 PM (9 years, 7 months ago) |
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ok. fucking 12 pages is ALOT to read man... but i said i would.
SO, to trouble shoot a jar issue, i would do the following.
1. Buy new jars. 2. Send me you addy so i can get you some SFD's. 3. 1/4" or 3/8" hole, drilled from the top so burs end up on the bottom. 4. RTV the sfd to the top.
Proceed how you do. Definitly do a test jar or two. If one of those test jars contams, then its the PC. (doubtful)
As stated earlier, im sure without a doubt that your jars are letting the contam in, SINCE, your test jars from earlier were contaming. Now, lets say you take your jars out hot, condensation forms under the foil due to difference in temp with outside, therefor wetting your filter allowing contams to get dragged in. The filters i will send you will be hydrophobic, meaning water forms drops on top of it rather than wetting the filter material. (mine are PTFE/Teflon)
i would try moving your SAB into a different room, wood tends to harbor more molds than one can imagine. Definitly be doing sterile work in a finished and painted room. Drywall and paint prevent most drafts from happening. Any draft will hinder your SAB sterility. If you have a small closet you can fit a small table and your sab in, go there. Clean it first obviously.
I know this is a bitch to trouble shoot because my last set of 26 jars i think i only got 5 or 6 good ones. Im sure its my sterile technique and my enviroment. I have black mold inside the walls of my apt, so it shows up in my dishes and jars. Kizzle has already ID'ed my contam as Asperillus Niger. Kinda wispy looking, white myc. Could be mistaken for cube, but this looks wispy and weak. when sporulating, it forms lil black balls, which powderize when disturbed. Thats aperillus niger, as far as i can tell. Pretty harmful to humans also i believe.
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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: TheEaglesGift]
#20383475 - 08/06/14 04:19 PM (9 years, 7 months ago) |
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Well I looked at everything small that I take for granted and the lid thing with my master is all I've come up with. Especially that could account for such a high failure rate.
I'd do even worse with PF cakes considering I've never done one nor read the tek on them. I've had plenty of successful grows and have 2 tubs now that are fine so I think going to a 1/2 pint cake from spawning 8qts at a time really would make me quit lol.
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blojo02184
Big Red
Registered: 05/15/13
Posts: 3,527
Loc: Maine
Last seen: 1 month, 17 days
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Re: Where are my contams coming from? [Re: Ghatti]
#20383493 - 08/06/14 04:23 PM (9 years, 7 months ago) |
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Quote:
Ghatti said: Well I looked at everything small that I take for granted and the lid thing with my master is all I've come up with. Especially that could account for such a high failure rate.
I'd do even worse with PF cakes considering I've never done one nor read the tek on them. I've had plenty of successful grows and have 2 tubs now that are fine so I think going to a 1/2 pint cake from spawning 8qts at a time really would make me quit lol.
i feel ya, once you get grains pf cakes are just childs play
but seriously, shoot me an addy and ill shoot out 6 or 12 disks. That way we can then rule out lids/jars and the PC.
Im guessing your jars, due to the fact that contams dont form in the grains but on top of everything.
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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: blojo02184]
#20383555 - 08/06/14 04:35 PM (9 years, 7 months ago) |
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Now I'm thinking that was a fluke or maybe those were jars that hadn't been cooked, or that that was just part of my problem.
The main thing is 99% of my jars colonize fine, I g2g them and those colonize fine, I then spawn them and my sub colonizes fine, only when I introduce fae do I find the nasty.
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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: Ghatti]
#20383563 - 08/06/14 04:36 PM (9 years, 7 months ago) |
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And I do appreciate the generous offer blojo but I ordered a set of 10 disks earlier that I should be able to make 40 out of.
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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: Ghatti]
#20383586 - 08/06/14 04:38 PM (9 years, 7 months ago) |
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Try prepping your jars as normal and sterilize a syringe with water. Go through the whole process as normal except inoculate with sterile water. Maybe inoculate 2 with sterile water, don't inoculate 2 at all, and open 2 for a few seconds in your SAB. Mark them all so you can tell what's what. Wait a week, or however long you normally see contamination.
The purpose is to test your method and filter without wasting costly spores. You could even inoculate with a blank agar wedge that has nothing growing on it.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
Edited by SpitballJedi (08/06/14 04:39 PM)
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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: SpitballJedi]
#20383603 - 08/06/14 04:42 PM (9 years, 7 months ago) |
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I need to run a set of controls indeed. I will leave one completely alone and the other I will go through the motions of cracking the lid and putting a fake wedge in tonight.
3 days ago just to test my sab I opened a petri for 15 secs to see what if anything was floating around maybe and so far the plate is still clean.
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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: Ghatti]
#20383607 - 08/06/14 04:43 PM (9 years, 7 months ago) |
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I really do think its because I've been sitting my master lid on a ring and recapping it at the least, 5 times. The most about 13
Edited by Ghatti (08/06/14 04:50 PM)
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blojo02184
Big Red
Registered: 05/15/13
Posts: 3,527
Loc: Maine
Last seen: 1 month, 17 days
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Re: Where are my contams coming from? [Re: Ghatti]
#20383886 - 08/06/14 05:26 PM (9 years, 7 months ago) |
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I tend to stay away from g2g because of the contamination risk.
If everything seems to colonize fine, and contams at the fruiting stage, then it is in your spawn. And that you cannot tell it apart from your culture.
My suggestion, is start a new one.
Since your using agar I am to assume this is not just a multi spore grow. During the time of transferring from plate to plate and I don't mean the motion of moving the agar through air but rather the act itself; I believe you are transferring contams... Along with perfect looking myc. It does happen I believe. Kind of like a symbiotic relationship.
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