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Offlinetryingtotrip
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Registered: 09/26/18
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Last seen: 6 years, 1 month
I'm getting mold 100% of the time (agar)
    #25523280 - 10/08/18 10:48 PM (6 years, 3 months ago)

I keep getting blue mold on all my agar slides. It starts off looking like myc and then the middle turns blue and then grows throughout the dish. I'll go over my procedure

-working with SAB in a wine cellar with temp at 63F and no air flow
-agar is MEA
-micropore tape for wrapping
-iso alcohol for sanitation
-wear a lab coat
-wash before procedure
-dishes are on a cookie cooling tray(above the boxes surface)

first I make agar and then sterilize with pressure cooker for 1 hour, then I let all the pressure leave the cooker then take jar over to the cleaned SAB and place inside with my petri dishes/sharpie/alcohol rag. spray lysol and let sit for 5 min. Then I pour my dishes and let them sit till solid. I inoculate them with a loop that is flame sterilzed and then once all of them are done I wrap them with micropore tape and label them and then close the SAB holes and then move box to the garage because it has a warmer temp 72F.


What mistake am I making to have all dishes be contaminated? I understand that you guys cant see me doing this procedure but I am trying really hard to be careful. Is there anything I can try to save the contaminated dishes? or dump them out and reuse the dishes if I wash them?


Thanks guys!

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InvisibleJHOVA
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25523288 - 10/08/18 10:53 PM (6 years, 3 months ago)

pics


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๐Ÿ…ƒ ๐Ÿ„ด ๐Ÿ„ฐ ๐Ÿ„ผ    ๐Ÿ„ฒ ๐Ÿ„ป ๐Ÿ„ธ ๐Ÿ„ฝ ๐Ÿ„ถ ๐Ÿ…† ๐Ÿ… ๐Ÿ„ฐ ๐Ÿ„ฟ

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OfflineDcevLearn
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25523290 - 10/08/18 10:55 PM (6 years, 3 months ago)

You don't need to pressure cook that long so it's probably not that.  Although you didn't mention venting the pressure cooker, so be aware of that.

I'm guessing you are inoculating the dishes from a spore print?  Do you only have one spore print to try?

More importantly, did you inoculate every dish or did you leave any alone?  If the dishes you left alone also become contaminated that will narrow it down.

Another thing to narrow down would be to leave the dishes inside the still air box after the transfer and not to mess around with trying to tape them.  Maybe you are contaminating the dishes through your handling at the end or your taping.


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Last Project: LAGM: Stones From Scratch in 2020


Edited by DcevLearn (10/08/18 10:56 PM)

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Offlinethe_way
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Re: I'm getting mold 100% of the time (agar) [Re: DcevLearn]
    #25523313 - 10/08/18 11:15 PM (6 years, 3 months ago)

could just be agents slipping in and contaminating your shit might try better locks


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They were dead before the ship sank!

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Invisiblevan hattonFacebookDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: JHOVA] * 2
    #25523320 - 10/08/18 11:19 PM (6 years, 3 months ago)

Quote:

JHOVA said:
pics



:begger:


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: the_way]
    #25523349 - 10/08/18 11:36 PM (6 years, 3 months ago)

I had 2 control plates that were not contaminated. the contamination follows the stream I made with the spores. I'll post pics tomorrow morning!

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OfflineDcevLearn
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25523714 - 10/09/18 06:23 AM (6 years, 3 months ago)

Did you make the print?  Just sounds like a dirty print so far.


--------------------

Last Project: LAGM: Stones From Scratch in 2020


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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: DcevLearn]
    #25523718 - 10/09/18 06:25 AM (6 years, 3 months ago)

Spray soapy water instead of Lysol. I use bleach+degreaser soap + water. Normal cleaning ratios of course, 1/10 bleach and a few drops of soap.

Edit: and wash ur hands and don't work over ur plates like she said ^^^ it's ok to get things wet with the soapy water too. It holds spores down like when it rains. Lysol is nothing but problems. It doesn't really "wet the air"

Edited by nube424 (10/09/18 06:28 AM)

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Offlinetrip_hazard
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Re: I'm getting mold 100% of the time (agar) [Re: nube424]
    #25523772 - 10/09/18 06:59 AM (6 years, 3 months ago)

:whathesaid:

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: trip_hazard]
    #25523887 - 10/09/18 08:32 AM (6 years, 3 months ago)

No I didn't make the print I bought it from someone.

Ok I will do soapy water instead of Lysol. I usually start off with Lysol and then i'll do a couple sprays of alcohol right before I get started.

I'll post some pictures in a couple minutes

thanks for the input guys. I tried a different print last night to try to eliminate if the print is dirty or not.

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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25523906 - 10/09/18 08:53 AM (6 years, 3 months ago)

A lot of prints are dirty. Even if u get mold u gotta separate the good mycelium from it as soon as u see mushroom mycelium.

Save contaminated ones for a few days in isolation. Sometimes spores germ a week after the first contamination.

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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: nube424]
    #25523909 - 10/09/18 08:58 AM (6 years, 3 months ago)

U could try making a small amount of spore solution and noccing up a little grain. Then grab a grain covered in mycelium and put that on agar. That's a decent way of getting mycelium from a dirty print. U can also inject spore solution into a brf puck and transfer mycelium to agar as soon as it appears.

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: nube424]
    #25523988 - 10/09/18 09:53 AM (6 years, 3 months ago)

dpnnt spray iso inside a sab. you will explode one day. describe a SAB in a wine cellar... pics bruh pics.

what kind of contamination? are your petri dishes wrapped and sterile before pouring?

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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524002 - 10/09/18 09:59 AM (6 years, 3 months ago)

It sounded like pennecilin. I know that's spelt wrong.

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524005 - 10/09/18 10:00 AM (6 years, 3 months ago)

Quote:

tryingtotrip said:
reuse the dishes if I wash them?





hard no.

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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524011 - 10/09/18 10:03 AM (6 years, 3 months ago)

Fun fact. They make shallow clear pp5 containers for restaurants. Those would be resuseable. Just as transparent as petris. I'll take a pic tomorrow when I go to work :wink:

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524013 - 10/09/18 10:04 AM (6 years, 3 months ago)

Here are some pics of the contam from day 3 to day 8 or 9

It's just a typical SAB from BOD's instructions and it's in a walk in wine cellar that has wine on the wall and then space to walk and I just set up a table and the SAB on the table and worked there.


Petri dishes are always brand new and I open then inside the SAB


Edited by tryingtotrip (10/09/18 10:08 AM)

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524018 - 10/09/18 10:06 AM (6 years, 3 months ago)

nube,
I think i'm going to order another print and I'll try your tips as well! thank you

Ok I wont reuse them!

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Invisiblenube424
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524022 - 10/09/18 10:07 AM (6 years, 3 months ago)

Is there a lot of exposed wood in there? Thats probly why ur having a problem. Try the soapy water tho.

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524025 - 10/09/18 10:08 AM (6 years, 3 months ago)

my first dishes were all trashed but i at least grew some myc. you got straight mold all around:sad:

you need new spores. try a shroomery sponsor.

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