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sgpa



Registered: 12/07/17
Posts: 24
Last seen: 2 years, 4 months
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WTF...what is this contam??
#26546964 - 03/20/20 03:15 PM (3 years, 10 months ago) |
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5 days or so after inoculating 9 grain jars with a GT clone (agar to grain), I am seeing this brown, coffee-grounds or dirt looking contaminant...what the hell is this? This is only my 3rd grow, and my previous jars all have done fine? I sterilized the jars @ 15psi for 90 min and used what I thought was good, clean technique when transferring the agar to the jars (SAB used, flamed the scalpel between transfers, etc)
I'm assuming these jars are toast, but would like some insight so as to learn from this experience!
Cheers, guys.
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,767
Loc: In the Gills
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Re: WTF...what is this contam?? [Re: sgpa]
#26546999 - 03/20/20 03:37 PM (3 years, 10 months ago) |
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Yeah that's mold it's toast sorry
Working in an SAB just doesn't get the perfection of a flowhood so you gotta do more attempts of the same thing's to increase your percentage of success.
Edited by rumfor69 (03/20/20 03:41 PM)
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sgpa



Registered: 12/07/17
Posts: 24
Last seen: 2 years, 4 months
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Re: WTF...what is this contam?? [Re: rumfor69]
#26547617 - 03/20/20 08:53 PM (3 years, 10 months ago) |
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Thanks for the feedback, but I want to figure out where/how this went south. Since 9/9 jars were affected, I would think it can’t just be “bad technique” with the agar transfer in the SAB, as some of the jars would be clean. I do recall thinking there seemed to be a little moisture on the inside of the jars after taking them out of the PC...could there have been too much water in the PC, thus boiling up over the top of the jars and getting water in through the filter disc? Would this even cause mold to form, if it happened?? I suppose I could have contaminated the agar plate when obtaining my piece of fruit tissue -this was my first attempt at cloning after all, so very possible. Please, any insight or advice would be much appreciated!
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antmanmax
Padawan



Registered: 03/22/19
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Re: WTF...what is this contam?? [Re: sgpa]
#26547667 - 03/20/20 09:24 PM (3 years, 10 months ago) |
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Most likely whatever mold hitched a ride on the agar. How many transfers did you go between the original clone plate and the plates you used for a2g?
Edited by antmanmax (03/20/20 09:25 PM)
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,767
Loc: In the Gills
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Re: WTF...what is this contam?? [Re: antmanmax]
#26547818 - 03/20/20 11:38 PM (3 years, 10 months ago) |
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Any moisture getting into the jars from the PC process is still sterilized so no mold came from there. Working with agar in an SAB instead of a flowhood is gonna be a dice roll. It's very easy for something to be in the still air and hitch a ride.
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oldest hippie
Stranger


Registered: 01/13/20
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Re: WTF...what is this contam?? [Re: sgpa]
#26547823 - 03/20/20 11:44 PM (3 years, 10 months ago) |
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Quote:
sgpa said: Thanks for the feedback, but I want to figure out where/how this went south. Since 9/9 jars were affected, I would think it can’t just be “bad technique” with the agar transfer in the SAB, as some of the jars would be clean. I do recall thinking there seemed to be a little moisture on the inside of the jars after taking them out of the PC...could there have been too much water in the PC, thus boiling up over the top of the jars and getting water in through the filter disc? Would this even cause mold to form, if it happened?? I suppose I could have contaminated the agar plate when obtaining my piece of fruit tissue -this was my first attempt at cloning after all, so very possible. Please, any insight or advice would be much appreciated!
I wonder if this might be ergot. Ergot infects rye grain and turns the grains black. If it is ergot, it is toxic and may even be fatal. LSD can be derived from some of the alkaloids it contains, but none for me please.
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hummingbird

Registered: 06/29/14
Posts: 2,134
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That looks like black pin mold (definitely not ergot), and the jars are toast. It either came from your agar wedge or the air when you inoculated. If that's your wedge its growing next to it probably came from your plate.
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sgpa



Registered: 12/07/17
Posts: 24
Last seen: 2 years, 4 months
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Re: WTF...what is this contam?? [Re: antmanmax]
#26548149 - 03/21/20 05:58 AM (3 years, 10 months ago) |
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I actually only did the one plating of the clone. I also have done several plates of my MS syringe, and they are looking very good. Perhaps I will use those for my next inoculation. I do have a couple questions regarding agar, since we are now into that subject... how long can you keep a plate viable, and if it has grown out to the edge of the plate, is that still good to use? I plan of sticking with agar, as it seems to be the ONLY way to go, from what I've read, but as stated, am a def noob here, so edumacate me!
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sgpa



Registered: 12/07/17
Posts: 24
Last seen: 2 years, 4 months
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Re: WTF...what is this contam?? [Re: hummingbird]
#26548203 - 03/21/20 06:40 AM (3 years, 10 months ago) |
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I agree it is probably not ergot, as this is not the grains themselves that have turned black, but a separate entity altogether...small pieces of dark brown, like dirt or fine coffee grounds. I am definitely tossing the contents of all jars, and starting from scratch, but as I've said, I would love to make this a learning lesson. BTW, I did find the plate I inoculated with the clone, and it had small dots throughout the agar as well, so I do believe I contaminated the plate when I initially transferred my clone piece to the plate!
Cheers.
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