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 Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk 
Last seen: 3 years, 18 days
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 GAH!!! 
#2400688 - 03/03/04 10:27 PM (20 years, 1 month ago) |
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Damn it... now two more of my 12 jars have contams. I don't understand why this shit is taking so long to colonize, it's been almost two weeks now and all i'm getting is tiny little splotches of myc. I followed the recipie to a T, and made it the best environment i could (80 temp for incubating). Yet 3 of my jars now have contams in them, one green, one black and one brown. I did get a little overzealous with the liquid innoc i put in, but I only did that to a few jars, and all of them seem to be fucking up royally. I just don't get it... any help or feedback, or sympathy  would be appreciated. 
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Will all the big boomers please unveil, please unveil, please unveil.
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 Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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 Re: GAH!!! [Re: Hrethic]
#2400732 - 03/03/04 10:40 PM (20 years, 1 month ago) |
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You should have posted in the contam forum, since the subject of your post is contamination.
Try again. Perhaps your inoculant was not sterile. What technique did you use to sterilize the jars? What technique did you use to filter the jars? What technique did you use to inoculate the jars?
Joshua
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The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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Anonymous
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 Re: GAH!!! [Re: Hrethic]
#2400737 - 03/03/04 10:42 PM (20 years, 1 month ago) |
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I think this belongs in basic cultivation.....as for your question..my guess is your liquid syringes were either contamed or you didn't sterlize the syringe properly or the air when innoculating.....or its possible you didn't pressure cook/steam long enough
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Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk
Last seen: 3 years, 18 days
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 Re: GAH!!! [Re: ]
#2400876 - 03/03/04 11:37 PM (20 years, 1 month ago) |
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I used glasses instead of 1/2 pint jars, i didn't have a pressure cooker, but that's why i presterilized the substrate. I'm using BRF jars and verm with tap water. I'm fairly sure that the innoc is sterile, i got it from ryche a few weeks ago and made a batch of Karo with it, which is still growing strong with 0% contam. How do you mean filtering the jars? I didn't read anything about filtering in the PF... that's another thing, i have now 3 teks at my disposal, all with conflicting reports as to what temperature and times things should be going at.
My own personal guess is that a large number of small bad things happened along the way that caused failure... i was hoping i'd get it my first time through. Ah well, that's what life is for... improvement. And sorry about the wrong forums, i didn't know quite which to post in because i'm talking about growing and contams and procedures...
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Will all the big boomers please unveil, please unveil, please unveil.
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Anonymous
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Re: GAH!!! [Re: Hrethic]
#2400886 - 03/03/04 11:42 PM (20 years, 1 month ago) |
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just keep trying and get some experience under your belt...you'll soon realize which teks are the real deal and which aren't and probably use different things from different teks till you find what works best and easiest for you....no one can really tell you what your problem is tho....if your sure your syringe is ok try some more jars/glasses
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Re: GAH!!! [Re: Hrethic]
#2400924 - 03/03/04 11:57 PM (20 years, 1 month ago) |
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Quote:
didn't have a pressure cooker, but that's why i presterilized the substrate.
Can you explain this? How did you get your pre-sterilized substrate into your glasses? What proceedure did you use to sterilize your glasses?
When I refer to filtration, I refer to the lid mechanism on your jars. Some people poke 1 hole, some several. Some cover with 1 layer of foil, some with two. What kind of cover did you use with your jars.
Joshua
--------------------
The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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mark2000
journeyman
Registered: 11/02/03
Posts: 61
Last seen: 11 years, 8 days
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Re: GAH!!! [Re: Joshua]
#2401259 - 03/04/04 02:03 AM (20 years, 1 month ago) |
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its ur temp
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Suntzu
Geek
Registered: 10/14/99
Posts: 1,396
Last seen: 14 days, 7 hours
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Re: GAH!!! [Re: Hrethic]
#2401884 - 03/04/04 09:12 AM (20 years, 1 month ago) |
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Quote:
Hrethic said:
Yet 3 of my jars now have contams in them, one green, one black and one brown.
IME multiple contaminants that include molds are likley not from poor innoculum. Molds and such are not probable survivors of the PC cycle [even a boiling cycle will take care of molds, usually]. One way or another, the substrate is getting hit with *primarily* airborne contaminants. As asked above, did you use a PC?
If a PC was used, and used properly, this kind of result can come from the cooling step. If you pop the stopcock and let the thing rapidly come down to atmospheric pressure, there's the possibility that airbornes can be introduced. IMO, PC's need to be cooled carefully and somewhat slowly, to allow whatever filter mechanism you have employed to work while all the pressures/temps are equilibrating. Once "properly" stabilized, jars are at a low risk from airborne contamination.
I don't think we can pinpoint the problem until we hear exactly how you prepared the substrate. Does 'pre-sterilizing' imply that you did PC?
The next step, other than possibly altering the sterilization procedure, would be to incorporate a negative control or two. These are extremely helpful in pinpointing contamination vectors. They are not a waste either, because if after a week or so of incubating you see nothing bad, you can innoculate at that point.
Hope things turn around for you. . .
*edit*--I just reread and saw that a PC was not used. All I can say is that it's pretty tough to get clean BRF/grain spawn without a PC. Keep trying!
Edited by Suntzu (03/04/04 09:14 AM)
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Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk
Last seen: 3 years, 18 days
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Re: GAH!!! [Re: Suntzu]
#2403505 - 03/04/04 04:12 PM (20 years, 1 month ago) |
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PC i'm assuming means a pressure cooker. I put the BRF and Verm in the oven at 315 degrees for 20 minutes before i made the jars, and then i boiled them without a pressure coooker in a regular pot. I did two layers of tin foil, and after i innocuted i didn't put them back on (as a fault of the tek i read). By the by, thanks for repsonding so quickly guys... i really do need just more experience in what works and what doesn't... and a clean fucking environment...
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Will all the big boomers please unveil, please unveil, please unveil.
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Re: GAH!!! [Re: Hrethic]
#2403549 - 03/04/04 04:28 PM (20 years, 1 month ago) |
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Baking the ingredients likely had very little effect on the contamination present.
If you didn't put the foil back on, does this mean the glasses were open to the air?
How long did you boil them for?
I used to stem/boil my jars. If done properly and long enough it can be an effective method of preparation.
Here is the tek I used when I started, it worked very well for me, other than the terrarium they suggest:
MMGG
Joshua
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The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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Zero7a1
Leaving YourWasteland
Registered: 10/23/02
Posts: 3,594
Loc: Passing Cloud
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Re: GAH!!! [Re: Hrethic]
#2404081 - 03/04/04 07:20 PM (20 years, 1 month ago) |
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Its important to remember what you do with your hands when your innoculating your jars.
Dont smoke weed or get fucked up before you do, or at least if you do, remember that you are handling some sensitive things  .
What i would reccommend getting is a whole bunch of alcohol wipe pads. Handy at your disposal... a clean surface to work on... so that means before you do any work... just spray it down with some cleaning chemical. There is also this air cleaning product called OUST... it says that it kills 99.9 % of airborne germs and the like... So spray down your room, dont leave the fan on. Let everything settle.. and begin your work. Work with gloves if you can... i just made sure my hands were clean.. and i did the oven tek + the whole oust in the air thing. just to be on the safe side. I wiped my needle down with the alcohol pad and flame sterilized, just to make sure no contams settled with the water. If you didnt prepare you syringe correctly or did not make a proper spore print. These could be other factors that harbor the mold... making it almost impossible for you not to get contams every time you innoculate.
How did you prepare your liquid cultures? Were they done very "cleanly"? Thats why i bought a pc... cause it just kept messing up. But i have since learned a lot about being clean... and how easily those contams can sneak past you!!!
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What?
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Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk
Last seen: 3 years, 18 days
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Re: GAH!!! [Re: Zero7a1]
#2404305 - 03/04/04 08:13 PM (20 years, 1 month ago) |
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I definitely see that cleanliness is much more important than i thought...and i thought it was very very important. I didn't wear gloves, but i did clean my hands and never smoked or got fucked up before doing it. The strange thing about my Karo set up is that it still is at 0% contamination, and i've been more than careless with it. I'm not proud to say that, of course, but now i at least know it's not the innoculation or karo that fucked it up. Thanks Joshua for the Tek, but that's actually the exact one i used, and i've been finding conflicting reports of temps and times etc... so i have no idea if that one is right or wrong, or basically if anything i'm reading is right or wrong, i just need to experiment more and find out what works for my situation.
I'm pretty sure now, going through some posts and reviewing in my head what exactly i did, that it's the strain that i got fucked over with. From ryche i ordered the Koh Samui Super Strain. I guess a lot of people on here have been having troubles, and the fact that i'm not in the cleanliest of places surely doesn't help.
Thank you all for your input, it's greatly appreciated... i guess now i get to sit back and either watch them all die off one by one or hopefully see some awesome mushroom power prevail...we shall see!
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Will all the big boomers please unveil, please unveil, please unveil.
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Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk
Last seen: 3 years, 18 days
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Re: GAH!!! [Re: Joshua]
#2404449 - 03/04/04 08:49 PM (20 years, 1 month ago) |
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Quote:
Joshua said:
Baking the ingredients likely had very little effect on the contamination present.
How can baking at 315 degrees for nearly a half hour have no effect if boiling doesn't even reach 212, and that's at sea level.
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Will all the big boomers please unveil, please unveil, please unveil.
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Joshua
Holoman
Registered: 10/27/98
Posts: 5,398
Loc: The Matrix
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Re: GAH!!! [Re: Hrethic]
#2404996 - 03/04/04 11:14 PM (20 years, 1 month ago) |
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Because as the water activity decreases so does the effectivity of heat sterilization. Water activity parallels moisture content. Spores can more easily survive high temperatures when their environment is dry, such as in flour and vermiculite even if mixed with water appropriate for a PF jar.
I don't think I'd trust 20 minutes. I sterilize on the safe side, I like to make sure all of the spores and bacteria die.
Joshua
--------------------
The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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Hrethic
A Human, Being
Registered: 01/05/04
Posts: 2,397
Loc: Passing through the bulk
Last seen: 3 years, 18 days
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Re: GAH!!! [Re: Joshua]
#2407169 - 03/08/04 01:05 PM (20 years, 1 month ago) |
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Damn Josh, you know your shit... thanks!
And as an update, it seems that with one more jar full of contams all the rest are doing fine and finally starting to grow a bunch of myc in different spots. Yippy! Thanks guys for the input, it's helped a bunch.
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Will all the big boomers please unveil, please unveil, please unveil.
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