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OfflineMolecularAltruist
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Need some advice.
    #26377814 - 12/12/19 08:47 PM (1 year, 2 months ago)

I've been trying to deal with agar. I initially was pouring the agar in half pint jars because that's all I had, but even though I was able to sterilize them and get spores to germinate I've found it difficult to isolate the mycelium from the few contams that made it inside. So I'm going to try to pour agar into some polypropelyne petri dishes I and then PC them.

My question is - Do I need to drill holes and put micro pore tape on the polypropelyne dishes before I PC them, or at all?

Also, I was wondering if, when the mycelium does sprout, if I could just isolate that wedge of agar and drop it in a LC? If a very small amount of contams followed into the LC, would that be a tremendous problem? Or could I just stick a syringe in there and only suck up whats clearly mycelium.

Any help is greatly appreciated.

I'm really tired of this learning curve. I just want some damn mushrooms to eat so I can better myself.


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Offlinenarmaduke
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Re: Need some advice. [Re: MolecularAltruist]
    #26377820 - 12/12/19 08:53 PM (1 year, 2 months ago)

Dishes do not need any modification, you just put the lid on and secure them shut with plastic wrap or parafilm. also you should do a couple transfers before you drop a wedge to LC or you're risking a contamd LC.


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OfflineMolecularAltruist
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Re: Need some advice. [Re: narmaduke]
    #26377841 - 12/12/19 09:04 PM (1 year, 2 months ago)

Okay thank you!


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Offlinenarmaduke
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Re: Need some advice. [Re: MolecularAltruist]
    #26377865 - 12/12/19 09:18 PM (1 year, 2 months ago)

you also might want to look over your sterile tek too if youre having trouble getting a clean plate doing it normally.


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OfflineMolecularAltruist
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Re: Need some advice. [Re: narmaduke]
    #26377927 - 12/12/19 09:52 PM (1 year, 2 months ago)

Quote:

narmaduke said:
you also might want to look over your sterile tek too if youre having trouble getting a clean plate doing it normally.




I'm mostly having troubles in my SAB. I can get clean agar to work on, but whenever I transfer spores to the agar there's always some contams getting in. Apparently having gloves installed and sealed onto the SAB is a bad idea. Apparently it creates a vaccum and when you move with the gloves it stirs up any contamination on the bottom of the SAB. So I'm going to try a different approach next go around.


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Offlineinsutama
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Re: Need some advice. [Re: MolecularAltruist]
    #26377938 - 12/12/19 09:55 PM (1 year, 2 months ago)

Ya ditch the glovebox you will have better success with a SAB


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OfflineMycologist217
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Re: Need some advice. [Re: insutama]
    #26377947 - 12/12/19 09:58 PM (1 year, 2 months ago)

Your spores always contaminate because spore syringes and prints are always dirty...period. whenever you are working with spores you should expect to have to clean the culture up.


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OfflineMycologist217
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Re: Need some advice. [Re: Mycologist217]
    #26377952 - 12/12/19 10:00 PM (1 year, 2 months ago)

don't over think the SAB...cut holes in a tote big enough to stick your arms through. Your done. Before our do work in SAB spray the inside with multi purpose cleaner with bleach. The large droplets will bat down any contaminates in the air and when settled you'll ha e as good environment as you can without Flowhood.


--------------------
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OfflineMycologist217
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Re: Need some advice. [Re: Mycologist217]
    #26377962 - 12/12/19 10:06 PM (1 year, 2 months ago)

When using your SAB sterile technique is the primary controller of vectors.

Proper sterile technique is a little tough to grasp but I've you understand it the movements become very intuitive.

Here's a quick start guide to good SAB sterile technique:

-Loosen jar lids or petri lids before attempting transfer.

-Rinse gloves with iso in-between each transfer

-sterilize tool with one hand while leaving the other hand frozen inside the SAB

-with free hand remove lid from transfer plate holding it above  dish cut wedge.

Place lid back on transfer dish, remove lid from transferee plate.

Make transfer

Replace lid...place tool aside

Tighten lid of fresh plate leaving mother plate loose...rinse repeat.


--------------------
My LC Manual (With custom LC lid Tek) ~~ Required Mycology Supplies ~~ Agar Work Videos ~~ L G M
AMU Q&A-NO SYMPATHY FOR THE DEVIL! KEEP THAT IN MIND! BUY THE TICKET: TAKE THE RIDE

Check out my Retail Gourmet Mushroom Farm!

Mycologist217 is a fictitious entity that uses images supplied by Google to mask his/her inability to develop normal social habits.


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