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OfflineBooooooooom
Stranger
Registered: 11/16/15
Posts: 139
Last seen: 6 months, 22 days
Agar contamination?
    #26439527 - 01/18/20 02:18 AM (4 years, 11 days ago)

Hello guys,
I need your expertise.
I made a sporesyringe from a relativly clean print. Then i made agar plates, 4% LME to 96% water and pressurecooked 25-30min.

Did everything in a SAB. But something doesnt look right, efter about 1week i see weird white circles, doesnt look like mycelium. Take a look:




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OfflineMrBovineJoni
Glip-Glop

Registered: 11/28/19
Posts: 264
Last seen: 13 days, 11 hours
Re: Agar contamination? [Re: Booooooooom]
    #26439598 - 01/18/20 03:54 AM (4 years, 11 days ago)

Difficult to tell exactly (pictures seem a bit blurry) and the no pour container plastic is also makes it more hazy so difficult to say for sure but that looks bacterial. Definitely does not looks like anything you should be seeing from a healthy MS inoculation after a week.

Quote:

I made a sporesyringe from a relativly clean print.



What do you mean by that? Was the print examined by a microscope? You would never be able to check that with a naked eye. Even if the print is done in sterile environment, the mushroom cap used for printing almost always carries contaminants.

Did you keep any of the no pour containers without inoculation as a reference sample? Now it is difficult to tell if your spores were carrying contaminants, or it was the no pour that was not clean or even the sterile technique in the SAB was not sufficient.


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InvisibleSonOfAHooker
chingchong
Registered: 03/11/19
Posts: 108
Re: Agar contamination? [Re: Booooooooom]
    #26439602 - 01/18/20 04:01 AM (4 years, 11 days ago)

Probably bacteria/yeast. Lower your mea closer to 2%. How much agar did you add?

Sometimes high water content contributes to faster contam rate. By the looks of it I'd guess you shot a bunch of liquid from the syringe instead of just 2-3 drops


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InvisibleLadysKnight
Hello Ladies
 User Gallery

Registered: 10/09/15
Posts: 1,670
Re: Agar contamination? [Re: SonOfAHooker]
    #26439779 - 01/18/20 07:24 AM (4 years, 11 days ago)

Try again without the syringe, it adds unnecessary vectors for contamination.


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OfflineBooooooooom
Stranger
Registered: 11/16/15
Posts: 139
Last seen: 6 months, 22 days
Re: Agar contamination? [Re: MrBovineJoni]
    #26440186 - 01/18/20 12:47 PM (4 years, 11 days ago)

Quote:

MrBovineJoni said:
Difficult to tell exactly (pictures seem a bit blurry) and the no pour container plastic is also makes it more hazy so difficult to say for sure but that looks bacterial. Definitely does not looks like anything you should be seeing from a healthy MS inoculation after a week.

Quote:

I made a sporesyringe from a relativly clean print.



What do you mean by that? Was the print examined by a microscope? You would never be able to check that with a naked eye. Even if the print is done in sterile environment, the mushroom cap used for printing almost always carries contaminants.

Did you keep any of the no pour containers without inoculation as a reference sample? Now it is difficult to tell if your spores were carrying contaminants, or it was the no pour that was not clean or even the sterile technique in the SAB was not sufficient.




hello,
yes i did keep some plates. I had rock hard LME that might have had a factor in the contamination. Becuase the plates are yellowish, it should look more lighter


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OfflineBooooooooom
Stranger
Registered: 11/16/15
Posts: 139
Last seen: 6 months, 22 days
Re: Agar contamination? [Re: SonOfAHooker]
    #26440188 - 01/18/20 12:48 PM (4 years, 11 days ago)

Quote:

SonOfAHooker said:
Probably bacteria/yeast. Lower your mea closer to 2%. How much agar did you add?

Sometimes high water content contributes to faster contam rate. By the looks of it I'd guess you shot a bunch of liquid from the syringe instead of just 2-3 drops




My agar was rock hard, i will have to buy new.


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