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Offlinestellabella11
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contams...any ideas where we went wrong?
    #7952778 - 01/29/08 08:51 PM (16 years, 3 days ago)

we did the pf tek. watched the video. followed directions to a T... sterilized the shit out of the room. steam sterilized the jars for 90mins. used a glove box. and we've thrown out 6 out of 12 jars so far. i can officially say that trich is the bain of my existence and i'm just not sure where we went wrong. the trich seems to be growing from the inoculation sites on the side of the jar? idk where we went wrong. we sterlized with the syringe after every jar.. i just dont get it :[ any ideas?


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I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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OfflineFrost000
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7952816 - 01/29/08 08:57 PM (16 years, 3 days ago)

Ideas.....CLEAN EVERYTHING. Take a shower before hand, wear surgical gloves, isopropyl alcohol EVERYTHING, spray lysol FIVE MINUTES beforehand. You cannot be a mushroom cultivator without proper sterile technique. Good luck next grow.


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InvisibleCrasher
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Re: contams...any ideas where we went wrong? [Re: Frost000]
    #7952872 - 01/29/08 09:05 PM (16 years, 3 days ago)

steam sterilized or pressure cooked?

How did you sterilize the needle after each jar?

Was there a filter layer between the lid and band (ie coffee filter)


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Give me silence, water, hope;
Give me struggle, iron, volcanoes...


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: Frost000]
    #7952876 - 01/29/08 09:06 PM (16 years, 3 days ago)

boo. we sterilzed to the point of sacrificing proper breathing. we lysol'd, isopropyl'd, showered, wore a friggen white suit slathered in alcohol... i'm obv sooo not even close to being a cultivator yet but where the hell did it go wrong :[ thanks for the response, btw


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I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: Crasher]
    #7952896 - 01/29/08 09:09 PM (16 years, 3 days ago)

steam sterilized- no pressure cooker.

flame sterilized. w/ an alcohol lamp.

and no filter...is that our problem?


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I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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Offlineaddy
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7952916 - 01/29/08 09:11 PM (16 years, 3 days ago)

Get a Pressure Cooker!! :dogpile:


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OfflineFrost000
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7952918 - 01/29/08 09:12 PM (16 years, 3 days ago)

Crasher had a great question how did you sterilize? Pressure cooker? A good possibility is that you have a contaminated syringe. Also, how are you incubating? Incubation above 75 degrees GREATLY increases the contamination probability. I've had NUMEROUS failure due to contamination(trichodermia in particular) but if you hone your sterile techniques you can overcome it.


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InvisibleCrasher
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Re: contams...any ideas where we went wrong? [Re: Frost000]
    #7952928 - 01/29/08 09:13 PM (16 years, 3 days ago)

Problem 1- No pressure cooker

Problem 2- flame and wipe syringe with alcohol

Problem 3- get some tyvek/polyfill/coffee filters to cover those holes.


--------------------
Give me silence, water, hope;
Give me struggle, iron, volcanoes...


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: Crasher]
    #7952976 - 01/29/08 09:19 PM (16 years, 3 days ago)

def investing in a pressure cooker for the next grow.  i had a feeling steam sterilizing wasnt going to do the job.  we did flame sterilize and wipe with alcohol. so i really think our sterilization practices after the jar sterilization were pretty great, but whats the use if the jars arent sterile?! so i guess we'll chalk this up to a learning experience and do better on grow 2? o and our incubator is set to 81? thought that was "ideal?"  :banghead:  :banghead:


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7952987 - 01/29/08 09:21 PM (16 years, 3 days ago)

thanks again for the help btw. :thanx: ..im sure these dumb mistakes get old to u guys real quick..


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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InvisibleCrasher
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7952988 - 01/29/08 09:21 PM (16 years, 3 days ago)

I incubated at 80 in a TiT because my grow area was always 55-65. If your room temp is in the 70's, don't worry about it.


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Give me silence, water, hope;
Give me struggle, iron, volcanoes...


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OfflineRogerRabbitM
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Re: contams...any ideas where we went wrong? [Re: Crasher]
    #7953195 - 01/29/08 09:55 PM (16 years, 3 days ago)

Hang on just a minute here. You don't have to use a pressure cooker with pf tek. You don't have to cover the holes with pf tek, other than with foil during steaming, which is removed after sterilization. You're supposed to flame sterilize between jars, but don't wipe with alcohol afterward. Simply use the needle a few seconds after you pull it from the torch. The first drop will cool the needle, and the rest will flow cleanly.

If the contaminant is growing from the injection sites, and you flame sterilized the needle, then your inoculant was contaminated. It isn't related to your sterilization procedure.
RR


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OfflineThe shroomy 1
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Re: contams...any ideas where we went wrong? [Re: Crasher]
    #7953220 - 01/29/08 09:59 PM (16 years, 3 days ago)

Guys, I think we missed the part where he says that the trich is at the inoculation points. If he was as sterile as he says then the problem could have been the syringe. He said that he didn't pressure cook. Something not necessary to the PF tek, (although recommended). If it was inproper sterilization of the substrate then it would have spread throughout at no specific point I'll give an example..

This is a pic of trich at a non specific area. Note the healthy mycelium growing from the inoculation point around the jar.

This pic clearly shows that trich began at dead center of inoculation point.

If he flame sterilized like he said... My money is on a bad syringe.

Just my .02


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AMU Q&A thread.


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: RogerRabbit]
    #7953236 - 01/29/08 10:00 PM (16 years, 3 days ago)

ok this may sound really stupid...but r u saying our syringes were contaminated? do i contact my supplier or did i do something to contaminate it? not really sure here... and thanks rr for replying to my post.. ur like the mushroom god. i feel like should curtsy or something <3


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7953260 - 01/29/08 10:02 PM (16 years, 3 days ago)

yeah, the trich is ONLY at the inoculation sites. not like spread out anywhere else in the jars. and im a "she" btw :]


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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InvisibleCrasher
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7953294 - 01/29/08 10:07 PM (16 years, 3 days ago)

Sorry, didn't mean to misinform anyone. I'm so used to PC'ing it becomes strange when others don't.

I always used filters over PF jars as well, just to be paranoid.

Agreed, nasty syringes.


--------------------
Give me silence, water, hope;
Give me struggle, iron, volcanoes...


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: Crasher]
    #7953313 - 01/29/08 10:12 PM (16 years, 3 days ago)

no worries crasher, def appreciate ur input regardless. does this kind of just happen? lol not sure if i have a syringe guarantee ir something..would be nice tho :]


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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OfflineThe shroomy 1
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7953335 - 01/29/08 10:17 PM (16 years, 3 days ago)

Quote:

stellabella11 said:
yeah, the trich is ONLY at the inoculation sites. not like spread out anywhere else in the jars. and I'm a "she" BTW :]




My sincere apologies.  :foreheadslap:  I truly meant no disrespect.

If you purchased from a sponsor on this site, then I'm sure they will have no problem making it right for you.


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AMU Q&A thread.


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InvisibleCrasher
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Re: contams...any ideas where we went wrong? [Re: stellabella11]
    #7953348 - 01/29/08 10:19 PM (16 years, 3 days ago)

Reputable vendors will attempt replacement given the circumstances.

Non-shroomery sponsors, however, are a different matter.


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Give me silence, water, hope;
Give me struggle, iron, volcanoes...


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Offlinestellabella11
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Re: contams...any ideas where we went wrong? [Re: The shroomy 1]
    #7953388 - 01/29/08 10:24 PM (16 years, 3 days ago)

hehe its all good. not many of my kind on the site. wellllll they USED to be a sponsor apparently, but are no longer.. kinda bummed. i just went to take look at the babies and ive noticed that the 6 i have left are all cambodian, so the other 6 that we tossed were all the golden teachers, so i def think it was the syringe now. there looks like some colonization on the remaining jars, so im keeping my fingers crossed they survive. green used to be one of my favorite colors until this week..


--------------------
I can't explain myself, I'm afraid, Sir, because I'm not myself you see..
-Alice


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