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Offlinetryingtotrip
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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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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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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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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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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.


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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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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524030 - 10/09/18 10:11 AM (6 years, 3 months ago)

Damn, alright thank you for the help guys. So if I get another spore print that is contaminated like this one is there anything I can do to make it work? Like using the sandwich method or anything like that?

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524034 - 10/09/18 10:12 AM (6 years, 3 months ago)

maybe try pf pucks. but mold is mold and dirty prints can be a real pain. i think in pic 3 you have some myc but its hard to tell.

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524044 - 10/09/18 10:18 AM (6 years, 3 months ago)

so if I get some growth in these dishes am I able to open then? and transfer to agar? Like If I open them they wont cause more problems with the spores spreading

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524059 - 10/09/18 10:27 AM (6 years, 3 months ago)

the point of agar is to clean your cultures. limit the time the contaminated dish is open. cut a healthy section and put to new petris.

but yours look like mine. which looks like i streaked with a mold swab:lol:

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25524258 - 10/09/18 11:59 AM (6 years, 3 months ago)

I feel like we got the same print bro lol

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OfflineKitchenChef
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25524403 - 10/09/18 01:08 PM (6 years, 3 months ago)

you live in an old mold infested straw house?

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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: KitchenChef]
    #25525064 - 10/09/18 05:49 PM (6 years, 3 months ago)

Quote:

KitchenChef said:
you live in an old mold infested straw house?





How'd you know?

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Offlinetryptkaloids
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25525142 - 10/09/18 06:23 PM (6 years, 3 months ago)

Just goes to show a picture is worth more than a thousand words. Your description made me think the micropore was just causing your culture to dry and bruise. But pics show that's defs mold. Use new spores as mentioned, and wrap those plates properly. Micropore is only good for a filter. You don't need to filter a petri dish


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
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OfflineVroomerMcZoomers
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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25525207 - 10/09/18 06:44 PM (6 years, 3 months ago)

OP Im going to come out on the side of bad print.  Sounds like you are doing it in a way that should work.

The only other thing I can think of for you to test it is setting up and mock inoculating some plates.  I mean, open the lid, swab with a flamed loop, everything but the spores.  See if your room is intolerably moldy somehow.

But I think it's your print.


--------------------
 

Either you repeat the same conventional doctrines everybody is saying, or else you say something true, and it will sound like it's from Neptune.  -Noam Chomsky

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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25525222 - 10/09/18 06:48 PM (6 years, 3 months ago)

Quote:

tryingtotrip said:
I feel like we got the same print bro lol




Those spores were clean. The contams i had were all bad technique related.

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Offlineukshroomer
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25525441 - 10/09/18 08:16 PM (6 years, 3 months ago)

My guess would be that it is the micropore tape. Someone please correct me if I am wrong. The right way to wrap dishes is with parafilm. If you cannot find it saran wrap works as well. I think it is the micropore tape. Not sure though so maybe a trusted cultivator can chime in.


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Offlineukshroomer
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Re: I'm getting mold 100% of the time (agar) [Re: ukshroomer]
    #25525457 - 10/09/18 08:21 PM (6 years, 3 months ago)

Actually now that you say the contamination showing on the dishes in the places where you streaked I am almost certain you have a dirty print. Get a good one from a vendor and start over.


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InvisiblemushboyMDiscord
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Re: I'm getting mold 100% of the time (agar) [Re: ukshroomer]
    #25525476 - 10/09/18 08:26 PM (6 years, 3 months ago)

micropore tape wrapping a petri isnt going to make or break contamination. using a single layer as a GE hole on a mini round might not be too cool tho.

its his print most definitely. id be on the horn with whoever sold that shit :helloyesthisisdouche:

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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25525503 - 10/09/18 08:35 PM (6 years, 3 months ago)

Based on my experience both personally and in micro labs, I would say your print is the source of contamination. Notice how the majority of the colonies are concentrated in the center of the dishes and not so much towards the edges? Either try a new print or work with dilution (wouldnt recommend dilutions, very costly and time consuming to test those).

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Offlinetryptkaloids
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25525655 - 10/09/18 09:28 PM (6 years, 3 months ago)

Quote:

mushboy said:
micropore tape wrapping a petri isnt going to make or break contamination. using a single layer as a GE hole on a mini round might not be too cool tho.

its his print most definitely. id be on the horn with whoever sold that shit :helloyesthisisdouche:



Definitely the print. But micropore isnt what id use to wrap a plate. :shrug: id ziplock em first


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25525668 - 10/09/18 09:32 PM (6 years, 3 months ago)

i just leave germ plates unwrapped sitting in the back of the sab. too lazy to wrap. ..bad practice?

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Offlinetryptkaloids
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Re: I'm getting mold 100% of the time (agar) [Re: mushboy]
    #25526315 - 10/10/18 08:15 AM (6 years, 3 months ago)

No, I do that with plates ready for grain. I transfer when it's ready then leave it unwrapped for a few days until i make grain. It should be fine as long as you dont have a fan running


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
Josex said:Don't take the site seriously bro, ain't worth it.
 

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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25526862 - 10/10/18 12:48 PM (6 years, 3 months ago)

i got contam on every plate for a while because i forgot to clean the SAB (wiping all sides with a sponge & soap + water) after pouring some agar.

Maybe you should do that just to be safe.


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Re: I'm getting mold 100% of the time (agar) [Re: l0wbob]
    #25526957 - 10/10/18 01:21 PM (6 years, 3 months ago)

So I grew out another set of the same spores and they all got mold again And my control which I just left them wide open with no lid got no contaminants in my SAB. I’m waiting on some other dishes that I did with some  spores from a vendor  to see if they got contaminated.  But if my controls didn’t get contaminated then it must be the spores right?

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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25527173 - 10/10/18 02:44 PM (6 years, 3 months ago)

It's never been your technique. Im not sure why you tried that print again...


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
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Josex said:Don't take the site seriously bro, ain't worth it.
 

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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25527552 - 10/10/18 04:49 PM (6 years, 3 months ago)

Just to make sure my technique is on par

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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25531463 - 10/12/18 05:59 AM (6 years, 3 months ago)

I would try what I said. Knock up some grain and transfer a grain that has mycelium on it to agar for further cleaning. Or brf puck if u don't have any grain.

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Re: I'm getting mold 100% of the time (agar) [Re: nube424]
    #25531534 - 10/12/18 06:52 AM (6 years, 3 months ago)

That's not going to work with a mold print.


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
Josex said:Don't take the site seriously bro, ain't worth it.
 

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Re: I'm getting mold 100% of the time (agar) [Re: nube424]
    #25531855 - 10/12/18 10:00 AM (6 years, 3 months ago)

ok i'll try out the grain to agar

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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25531887 - 10/12/18 10:08 AM (6 years, 3 months ago)

I wouldn't...


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
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Josex said:Don't take the site seriously bro, ain't worth it.
 

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Offlinetryptkaloids
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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25531889 - 10/12/18 10:08 AM (6 years, 3 months ago)

The whole point of agar is so you dont waste time and materials prepping grain


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"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25532079 - 10/12/18 11:37 AM (6 years, 3 months ago)

ok I wont then haha I got another print so I'll test it out on agar.

so I kept those dishes around and this is what's growing now. Do you guys think this is mold or pan myc? A few of them have these white growths with no sign of blue mold.


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Offlinetryptkaloids
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25532290 - 10/12/18 01:15 PM (6 years, 3 months ago)

I would transfer it and find out


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
Josex said:Don't take the site seriously bro, ain't worth it.
 

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InvisibleJHOVA
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Re: I'm getting mold 100% of the time (agar) [Re: tryptkaloids]
    #25532342 - 10/12/18 01:41 PM (6 years, 3 months ago)

Take a piece now thats furthest away from the mold.


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Offlinetryingtotrip
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Re: I'm getting mold 100% of the time (agar) [Re: JHOVA]
    #25532527 - 10/12/18 02:52 PM (6 years, 3 months ago)

sounds good :thumbup:

may not have time today but I'll get to it early tomorrow morning

Thanks guys I really appreciate it

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Offlinemaui
Registered: 11/20/14
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Re: I'm getting mold 100% of the time (agar) [Re: tryingtotrip]
    #25626552 - 11/19/18 11:46 PM (6 years, 1 month ago)

That, my frand, is once again... mold.

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