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Offlineplotikai
Stranger
Registered: 04/21/20
Posts: 23
Last seen: 3 years, 4 months
Cant get agar right
    #26850647 - 07/28/20 08:00 PM (3 years, 6 months ago)

Why can't I get proper germination on agar plates, I've used different syringes with different providers... This just tells me that I'm the problem and doing something wrong...

- My plates come from a sterile bag
- into a cleaned SAB
- I PC my agar at 15psi for 90 minutes
- let it cool down enough to handle (I get condensation buildup, is this what's causing the contamination??)
- put the plates back into the bag and leave overnight to cool
- I flame sterilize my needle
- place 1 drop in each plate
- rub with a flame sterilized inoculation loop
- wrap in parafilm

As you can see from the pictures, this looks exactly like my last failed attempt, nothing but bacteria? But why is it only in the spot I put the drop? wouldn't there be spots growing all over? Why specifically the drop that I placed the spores?



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Invisiblerickyswamps
Bad Apple
I'm a teapot


Registered: 11/08/18
Posts: 1,192
Re: Cant get agar right [Re: plotikai]
    #26850668 - 07/28/20 08:08 PM (3 years, 6 months ago)

More than likely your syringe is contaminated.


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OfflineGan
Wielder of Narya
Male


Registered: 08/26/19
Posts: 927
Loc: Valinor
Last seen: 6 months, 15 days
Re: Cant get agar right [Re: rickyswamps]
    #26850676 - 07/28/20 08:14 PM (3 years, 6 months ago)

Whoa you PC it for 90 minutes? 20-25 should be good, depending on how much sediment you have from your recipe.

It is strange, as I would expect the bacteria to follow your streak pattern a little more closely. Are you putting the drop in the middle and streaking it or are you putting it towards an edge and streaking over the entirety of the plate?

Also, spore prints tend to be easier to deal with on agar. I assume you've probably spent more $$$ than you'd like on syringes from different vendors, so head over to the Marketplace and I'm sure someone will send you a print no charge to get you started. I would but I dont have many. If I find any extra I'll PM you and mail it to you.


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OfflineMycochopper
Stranger than you
 User Gallery

Registered: 07/15/20
Posts: 82
Last seen: 1 year, 21 days
Re: Cant get agar right [Re: rickyswamps]
    #26850690 - 07/28/20 08:21 PM (3 years, 6 months ago)

I actually use catchup cups with agar in a SAB because its cost effective and I can afford to loose a couple.  I stack them 3 high and a fourth on top with almost boiling water and let them cool that way.  Im guessing that through thermodynamics the hot water on top helps prevent condensation in the agar cups and I get far less contams in my agar cups. Also I use a loop to spread a single drop of spores on the agar.


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OfflineMartinsapin
no-mad
Male User Gallery


Registered: 09/12/16
Posts: 310
Loc: Canada Flag
Last seen: 1 year, 8 months
Re: Cant get agar right [Re: Mycochopper]
    #26850719 - 07/28/20 08:34 PM (3 years, 6 months ago)

PC it for 45 min.
Ive never liked Spore syringe to agar.

PF tek then print then agar, grains,tubs. Or clone then agar. You ll be started for real


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looking for a sclerotia producer print


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