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Invisiblechimpezman
Stranger Danger


Registered: 03/21/13
Posts: 40
Loc: USA Flag
Agar Troubles [w/ pics]
    #19051237 - 10/29/13 09:59 AM (10 years, 3 months ago)

Hey all,

I started Agar a few days ago. It has been ~5 days since inoculation. I used a PDYA recipe that Frank has used in his agar experiments and noted was most promising. I used an inoculation loop (pretty awful technique as you can see from the smear patterns lol) from some prints I had. And One of them with a syringe.

My trouble is, they don't appear to be taking hold at all. Maybe I'm just impatient? After noticing this thread:HERE The suggestions there were to toss it all. Should I start over and do the same?

I have attached pics below:


The yellow/orange spots are the yeast. I definitely need to agitate the agar much better next time. I got a lot of clumping. Anywho...suggestions? Should I just use a syringe the next time? Or continue with inoculation loop?

I made about 12 dishes, and only 2-3 have progressed to show ANYTHING, and here are 2 of them. The rest have visible black spores on them, but nothing to show for it. :sad:

Thanks


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Invisible8ow8
Soylent McGriddle
Male


Registered: 05/22/13
Posts: 196
Loc: Canada
Re: Agar Troubles [w/ pics] [Re: chimpezman]
    #19051356 - 10/29/13 10:32 AM (10 years, 3 months ago)

Patience!

If you started from a dry spore print, you can expect up to two weeks before you see visible growth. I've used agar literally hundreds of times and only see growth that fast with pre-hydrated spores (i.e., from a syringe). They have to hydrate before they start growing, and that process adds a few days to the process....

I've never been able to get yeast nutrient to dissolve completely, lol.

If you don't see serious contams, there's no reason to throw them out. Even if you do have slimy, bacterial blobs, you *might* get lucky and see enough growth nevertheless that you can isolate some clean mycelium.

Here are some possibilities for serious issues with agar:

- The sugar content is more than 4%
- The incubation temperature is way off (depends on what you're growing)


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Edited by 8ow8 (10/29/13 10:35 AM)


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Invisiblechimpezman
Stranger Danger


Registered: 03/21/13
Posts: 40
Loc: USA Flag
Re: Agar Troubles [w/ pics] [Re: 8ow8]
    #19051450 - 10/29/13 10:58 AM (10 years, 3 months ago)

Duly noted on hydrating spores before agar! I was hoping to cut out steps by not using syringes but if its faster, that's important to note. I appreciate the advice. I'll definitely wait it out and see where things go. :thumbup:


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OfflineOUboundMycologist
Student Pharmacist
Male

Registered: 09/15/13
Posts: 49
Loc: USA Flag
Last seen: 9 years, 6 months
Re: Agar Troubles [w/ pics] [Re: chimpezman]
    #19051696 - 10/29/13 12:01 PM (10 years, 3 months ago)

Also if you use syringes on agar plates you only need (1 DROP).  Water is a source of contaminates and you want as little water on the agar as possible.  I use Y-Plates (3-sections) and the corner of each section I put one drop. (So 2 drops in each corner of each section)  Should see growth within 3-5 days.


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"Be the change that you want to see in the world"

Anything illegal or that has to do with mushrooms that I talk about is completely 100% fictional and is not true.


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