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OfflineBobBee
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Spore print to agar: how?
    #26405829 - 12/29/19 07:32 AM (4 years, 1 month ago)

If contaminants grow generally quicker than spore germination, how does one use agar to grow mycelium from wild spores or spores collected from my own mushrooms? Is the only solution to prepare so many agar plates that one eventually won’t have contamination on it? I know my spores have contamination because I did not collect them in a sterile environment. So, how do I use those spores to grow more mushrooms? Do I leave contaminated plates to grow for a couple weeks and see what happens?


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Invisiblemu5h13
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Re: Spore print to agar: how? [Re: BobBee]
    #26405842 - 12/29/19 07:48 AM (4 years, 1 month ago)

Suck some boiling water into a syringe then leave it to cool (with cap on)

In an SAB:
-Take clean shotglass and thoroughly wipe with IPA then let it dry
-Scrape spore print into shot glass
-Squirt cooled water into shot glass with spores
-Suck back up into syringe (can repeat the squirt and suck if spores are clumping together)
-Then leave the syringe for a few days to allow the spores to hydrate

-Then take your agar plate and, after shaking the syringe, drop the TEENIEST drop of liquid onto the plate. (If you have an sterile inoculation loop or suchlike you can smear the liquid a bit on the plate to spread it out, but I've had plenty of success just tilting the plate to make the drop run around)

Then leave it to incubate and see what you get

To isolate you want to find a plate that has a bit of distance between any growing contams and the mycelium and take a small chunk of mycelium and agar from a part furthest from the contams, which you then transfer to new agar and keep going until no contams.


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Invisiblemu5h13
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Re: Spore print to agar: how? [Re: BobBee]
    #26405846 - 12/29/19 07:49 AM (4 years, 1 month ago)

But yeah, try a few things and see what works best. Try growing some out, try isolating early etc..

Do that science thing.


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OfflineSynKyd
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Re: Spore print to agar: how? [Re: mu5h13]
    #26405899 - 12/29/19 08:24 AM (4 years, 1 month ago)

You can just scrape the dry print over the agar using typical SAB and good clean processes, a moist mix in the agar works best for this method.  Get a few plates going and expect to transfer several times to outrun contams in either method.


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Edited by SynKyd (12/29/19 09:20 AM)


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OfflineMariochi64
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Re: Spore print to agar: how? [Re: SynKyd]
    #26406019 - 12/29/19 09:55 AM (4 years, 1 month ago)

Buy a bacteria inoculating loop and a oil wick lamp apply some heat to the wire and then sterilize it with some rubbing alcohol, then take a small swipe of agar from your plate (do this in a still air box) then swipe some spores of your print and reapply it to the plate.( I COULD BE WRONG THO THIS COULD BE A VERY CRAPPY METHOD)


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Edited by Mariochi64 (12/29/19 09:56 AM)


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Invisiblemu5h13
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Re: Spore print to agar: how? [Re: SynKyd]
    #26406087 - 12/29/19 10:37 AM (4 years, 1 month ago)

Quote:

SinysterKyd said:
You can just scrape the dry print over the agar using typical SAB and good clean processes, a moist mix in the agar works best for this method.  Get a few plates going and expect to transfer several times to outrun contams in either method.




Ahh yeah i've never tried it that way, but i like to have my spores hydrated first, I've always reasoned that it helps give them a slight headstart against contams lol

But maybe it doesn't work like that at all, it's just how I do it :crazy2:


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InvisibleBph
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Re: Spore print to agar: how? [Re: mu5h13]
    #26406248 - 12/29/19 12:06 PM (4 years, 1 month ago)

I use sterilized q tips to streak spores. And the idea is things grow on the agar together and you pick the colonies you want and leave the others behind.


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OfflineAsuraS
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Re: Spore print to agar: how? [Re: Bph]
    #26406263 - 12/29/19 12:15 PM (4 years, 1 month ago)

Quote:

Bph said:
I use sterilized q tips to streak spores. And the idea is things grow on the agar together and you pick the colonies you want and leave the others behind.




This is what I do as well

https://www.shroomery.org/forums/showflat.php/Number/26325679


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InvisiblePsicomb
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Re: Spore print to agar: how? [Re: BobBee]
    #26406326 - 12/29/19 01:00 PM (4 years, 1 month ago)

I take one plate and I use it specifically for cooling my knife after flame sterilizing and then I cut a small wedge out of that plate, rub it on a spore print, and then use it to inoculate another clean, sterile agar plate. 

Repeat as many times as you need using the same cooling plate and the same print. 

This technique has worked well for me for hundreds of plates.  Some of your plates may have contams in them because of being wild prints but just keep transferring from the cleanest spots and you'll be good. Make a bunch of plates initially too.


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When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Edited by Psicomb (12/29/19 01:02 PM)


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OfflineDoyledozo
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Re: Spore print to agar: how? [Re: Psicomb]
    #26406532 - 12/29/19 03:18 PM (4 years, 1 month ago)

Thats a great idea!


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OfflineAsuraS
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Re: Spore print to agar: how? [Re: Doyledozo]
    #26406579 - 12/29/19 03:48 PM (4 years, 1 month ago)

I am really not seeing how this is a good idea, but maybe I am missing something.

With multiple streak plates, you are spreading everything out and (imo) have a better
chance of getting nice colonies.


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InvisiblePsicomb
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Re: Spore print to agar: how? [Re: Asura]
    #26406581 - 12/29/19 03:51 PM (4 years, 1 month ago)

I've never had luck with swabs myself:shrug: I've tried soft agar, hard agar, etc and more often than not it won't germinate at all.  I know others, such as you asura, are luckier with swabs.  I dont see what's really wrong with my method though.  You can use that piece of agar to streak a bit as well / break up some to create multiple inoculation points.  Just a lil creativity and it streaks perfectly fine.


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When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Edited by Psicomb (12/29/19 03:55 PM)


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OfflineAsuraS
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Re: Spore print to agar: how? [Re: Psicomb]
    #26406601 - 12/29/19 04:07 PM (4 years, 1 month ago)

Ha yeah, good point, Psi. I knew I was missing something - streaking the plate with the agar.

This might pretty good for those hard to germ prints. I have a few so I'll give it a try.


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OfflineDoyledozo
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Re: Spore print to agar: how? [Re: Psicomb]
    #26406603 - 12/29/19 04:09 PM (4 years, 1 month ago)

Maybe im just being impatient and a little overly worried. Put spores on plates 10 days ago. A few were swabbed and a few were done with a loop. No sign of life on any of them. Its my first time with anything other than a syringe. I should just be more patient.


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InvisiblePsicomb
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Re: Spore print to agar: how? [Re: Asura]
    #26406638 - 12/29/19 04:49 PM (4 years, 1 month ago)

Glad it sounds good to ya :laugh: I'll need to try your method too with using a sterile swab on a print.  Every time I've tried using a swab it was one I've bought / traded for, not one I added the spores to myself using a print, so having it fresh on the swab before streaking may be my ticket as well

Doyle....yeah 10 days is a bit but I'd still give it another week before tossing. Fingers crossed and a few prayers to baby jebus and hopefully you'll see some germination hah


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When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Edited by Psicomb (12/29/19 04:52 PM)


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OfflineDoyledozo
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Re: Spore print to agar: how? [Re: Psicomb]
    #26406645 - 12/29/19 04:59 PM (4 years, 1 month ago)

Do spores tend to eat little holes in the surface of the agar before the start to grow? Definitely seeing alot of that in my streak lines. Ill feel alot better once i see some white on one or more of them. Slam dunked one of my swabs last night to hedge my bet.


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Invisiblestaytrippy420
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Re: Spore print to agar: how? [Re: Doyledozo]
    #26406956 - 12/29/19 09:44 PM (4 years, 30 days ago)

I believe spores to plates works best with fresh spores. Some varieties are faster than others.
I've got some aggressive LGT plates making my PM look hella slow and difficult.

I've used both swabs and inoculation loops and I prefer swabs for doing more then one plate at a time.


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