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Offlineplutoniusspasticus
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Petri dishes keep contaminating
    #26886204 - 08/18/20 07:27 AM (3 years, 5 months ago)

I'm trying improve my sterile technique so I made 20 agar plates. I checked them 5 days later and they all have tiny white blobs on them. I used a sab, misted it with soapy water, used slow movements, minimized exposure time when opening the petris, didn't let my hands go over the petri dish. I pced the agar for 45 minutes and let it cool for 2 hours. I took a shower before. I used plenty of iso to clean everything. I used gloves and regularly cleaned them with iso. I work in a small room without air movement.

Here's a picture of one:


I don't know where the contam is coming from. Any advice would be great.


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InvisibleLadysKnight
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26886287 - 08/18/20 08:39 AM (3 years, 5 months ago)

I used to get those when I misted my sab.


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OfflinePrincessRosalina
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Re: Petri dishes keep contaminating [Re: LadysKnight]
    #26886389 - 08/18/20 10:00 AM (3 years, 5 months ago)

Quote:

LadysKnight said:
I used to get those when I misted my sab.




Do you just not mist anymore?


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InvisibleLadysKnight
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Re: Petri dishes keep contaminating [Re: PrincessRosalina]
    #26886406 - 08/18/20 10:09 AM (3 years, 5 months ago)

Correct.


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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: LadysKnight]
    #26886415 - 08/18/20 10:18 AM (3 years, 5 months ago)

It seems strange that spraying would be the problem. I'll try not spraying anyway.

Any other ideas anyone?


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Invisiblemaxmush
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26886436 - 08/18/20 10:27 AM (3 years, 5 months ago)

Hmmm. Are those specks growing at all?


--------------------
Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.


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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: maxmush]
    #26886476 - 08/18/20 10:41 AM (3 years, 5 months ago)

I've only just seen them so I don't know yet tbh. Up-close they look like white droplets.


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OfflineA.k.aM
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26886678 - 08/18/20 12:41 PM (3 years, 5 months ago)

It does look like bacteria but it might just be sediment it’s hard to say for sure without seeing them in person. Are they in the agar or on the surface?

Have you opened them since you made them or did you just pour them and wrap them?


--------------------
LAGM2020


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Offlinetryptkaloids
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Re: Petri dishes keep contaminating [Re: A.k.a]
    #26886717 - 08/18/20 12:55 PM (3 years, 5 months ago)

Don't blame misting entirely. Blame overmisting. You dont need to soak the top


--------------------
"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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Pick a book, Make some chips!
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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: A.k.a]
    #26886738 - 08/18/20 01:05 PM (3 years, 5 months ago)

@A.k.a
They're on the surface. I just poured them and wrapped them, haven't opened them.


Edited by plutoniusspasticus (08/18/20 01:09 PM)


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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: tryptkaloids]
    #26886742 - 08/18/20 01:07 PM (3 years, 5 months ago)

@Onlinetryptkaloids
I used a fine mister and I remember doing 3 sprays. Maybe it's still too much though.


Edited by plutoniusspasticus (08/18/20 01:08 PM)


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Offlinetryptkaloids
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26887179 - 08/18/20 05:41 PM (3 years, 5 months ago)

Did you wipe your agar vessel before you poured?


--------------------
"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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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: tryptkaloids]
    #26888084 - 08/19/20 09:58 AM (3 years, 5 months ago)

Yep, I cleaned the petri sleeve and the agar bottle with iso when I put them in the sab.


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Offlinetryptkaloids
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26888456 - 08/19/20 01:38 PM (3 years, 5 months ago)

What was your media? Sterilization times? Did you remember to vent your PC.


--------------------
"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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OfflineSpore211215
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Re: Petri dishes keep contaminating [Re: tryptkaloids]
    #26888495 - 08/19/20 02:04 PM (3 years, 5 months ago)

Could be what you’re misting with has contams hiding inside it. Are you using Iso, soapy water, or both? Maybe if the water is old it could be bringing stuff in you don’t want. I used to have a lot of problems with that but I tuned in my misting and hand movements and now I only get contams when I’m being careless


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Offlinetryptkaloids
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Re: Petri dishes keep contaminating [Re: Spore211215]
    #26888537 - 08/19/20 02:22 PM (3 years, 5 months ago)

Could also be you were bumping the box. Did you rest your arms on the holes?


--------------------
"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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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: tryptkaloids]
    #26888780 - 08/19/20 04:35 PM (3 years, 5 months ago)

I used lme. 45 minutes pced then I leave it in there for 2 hours to cool down. I used to instantly open once the pressure valve went down- that caused a lot of contamination.


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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: Spore211215]
    #26888794 - 08/19/20 04:37 PM (3 years, 5 months ago)

I use a new batch of purified water and soap for each session. I didn't wait long for the mist to settle though. I saw a post from bod or RR and they said to work immediately.
I wipe the sab and whatever I'm working with before I start. Perhaps I'm a little rough sometimes with my movements but I wouldn't think it would cause them all to contaminate.


Edited by plutoniusspasticus (08/19/20 04:52 PM)


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Offlineplutoniusspasticus
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Re: Petri dishes keep contaminating [Re: tryptkaloids]
    #26888801 - 08/19/20 04:40 PM (3 years, 5 months ago)

I may touch the arm holes but I don't rest on them. I don't think I bumped it last time.


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Offlinetryptkaloids
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Re: Petri dishes keep contaminating [Re: plutoniusspasticus]
    #26888841 - 08/19/20 04:59 PM (3 years, 5 months ago)

Are you letting your PC vent steam for 10 min before putting the weight on?


--------------------
"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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