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OfflineTheCyndicate
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Spore syringe and agar.
    #24018167 - 01/17/17 06:46 PM (7 years, 13 days ago)

All you TC out their, what is your go too agar recipe for spore germination when using a spore syringe?

Cyn


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InvisibleAdden
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Re: Spore syringe and agar. [Re: TheCyndicate]
    #24018175 - 01/17/17 06:48 PM (7 years, 13 days ago)

I'm not a TC but use 7g agar, 1.5 cups water, drop of Karo and pinch of potato flakes. I think that's pretty standard. Also it's easier to put spore solution on a sterile swab. You get a better streak and more predictable plates.


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InvisiblemushboyMDiscord
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Re: Spore syringe and agar. [Re: Adden]
    #24018249 - 01/17/17 07:13 PM (7 years, 13 days ago)

i use standard pasty agar.

quickest was 3 days. longest was 12-13.. it has germinate multiple cubes and 2 pans. good stuff.


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Offlinethemadextractor
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Re: Spore syringe and agar. [Re: mushboy]
    #24018314 - 01/17/17 07:38 PM (7 years, 13 days ago)

I use standard recipe. If I have trouble I will do a pasty plate half full of grain and then transfer grains to agar for cleaning.


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OfflineDeadPhan
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Re: Spore syringe and agar. [Re: themadextractor]
    #24018559 - 01/17/17 09:15 PM (7 years, 13 days ago)

Quote:

themadextractor said:
I use standard recipe. If I have trouble I will do a pasty plate half full of grain and then transfer grains to agar for cleaning.




I've had alot of trouble with syringe to agar skme reason. Not as much syringe to grain. My agar is great for transfers, clones, spore print, or colonized grain. I don't know why but I always have issues with germinating from spore syringe straght to agar. But I do agar in half pint wide masons so that's a good idea to inoculate a half pint jar that was filled with grain and then go from there. Right on.


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Offlinesynthetik
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Re: Spore syringe and agar. [Re: Adden]
    #24022791 - 01/19/17 12:10 PM (7 years, 11 days ago)

Quote:

Adden said:
I'm not a TC but use 7g agar, 1.5 cups water, drop of Karo and pinch of potato flakes. I think that's pretty standard. Also it's easier to put spore solution on a sterile swab. You get a better streak and more predictable plates.




wish I read this before I noc'd my plates up with a gt syringe, I over shot a drop... :smile: now all of them have a white liquid, not sure what that is...


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InvisibleMycolorado
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Re: Spore syringe and agar. [Re: synthetik]
    #24022806 - 01/19/17 12:17 PM (7 years, 11 days ago)

It's bacteria.  Using a swab would change nothing.


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InvisiblebodhisattaMDiscordReddit
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Re: Spore syringe and agar. [Re: Mycolorado]
    #24022827 - 01/19/17 12:27 PM (7 years, 11 days ago)

If you have syringe with bacteria making a streak plate will help dramatically.


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InvisibleMycolorado
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24022828 - 01/19/17 12:28 PM (7 years, 11 days ago)

It's not going to change the fact that it's contaminated...


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Re: Spore syringe and agar. [Re: Mycolorado]
    #24022831 - 01/19/17 12:33 PM (7 years, 11 days ago)

So :shrug: that's not the point. If we could decontaminate syringes we wouldn't have to worry about proper plating techniques


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Re: Spore syringe and agar. [Re: bodhisatta]
    #24022840 - 01/19/17 12:35 PM (7 years, 11 days ago)

It is the point...unless my reading comprehension is wrong (totally possible) OP made it sound like he wouldn't have milky buildup if he used the swab.  Also, there's no reason to use a tool when you can simply roll the bead of water around the plate, effectively streaking the spores. :shrug:


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Re: Spore syringe and agar. [Re: Mycolorado]
    #24022842 - 01/19/17 12:38 PM (7 years, 11 days ago)

Rolling a bead of water around is absolutely no substitute for a streak plate made with an inoculation loop. Not even close. A drop rolled around is fail if the syringe isn't clean. A streak plate is how to avoid the fail.


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InvisibleMycolorado
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24022844 - 01/19/17 12:39 PM (7 years, 11 days ago)

Don't worry dude, you might hit TC at 30K posts...


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Re: Spore syringe and agar. [Re: bodhisatta]
    #24022855 - 01/19/17 12:44 PM (7 years, 11 days ago)








one drop rolled around

this would be fail if there was bacteria



compare with properly plated spores




both spread plates, obtaining good mycelium on one of them, as you streak you dilute contamination which is in a lower concentration than the intended spores.

another few streak plates




if you have a syringe that has
100,000,000 spores
and
1,000,000 bacteria

if you spread it out eventually you can weed out bacteria since it's a 100:1 ratio of good to bad

that's a theoretical example but if you have any contaminants in a syringe and you just put a drop on agar and roll it around, you'll have a bad time

if you have a contaminated syringe and you don't try a proper streak plate it's operator error since you didn't try proper technique before giving up


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Offlineenlightenment
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Re: Spore syringe and agar. [Re: Mycolorado]
    #24022862 - 01/19/17 12:46 PM (7 years, 11 days ago)

Come on guys... chill

This is how to inoculate with a loop:

I make it a little different and rotate just 2 times

If the syringe is bacterial contaminated it's still hard to isolate healthy sectors especially for a novice. But it might be worth a try.


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Re: Spore syringe and agar. [Re: enlightenment] * 1
    #24022870 - 01/19/17 12:48 PM (7 years, 11 days ago)

that GIF is more for bacteriological work, for yeast and fungi(spores) you streak the whole plate and don't need to flame and then do more zigzags


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Offlinelovelaughlibs
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Re: Spore syringe and agar. [Re: enlightenment]
    #24022874 - 01/19/17 12:51 PM (7 years, 11 days ago)

Cheers bodhi, the pics made me 100% sure the plates i got rid of were bacterial after all.


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Offlineenlightenment
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24022878 - 01/19/17 12:52 PM (7 years, 11 days ago)

Thats why I wrote I make it a little different. It was meant as an illustration to get an idea about streaking.

It worked well so far. But if the syringe is contaminated as hell you don't have a chance.
I wasted over 20 petri dishes for a mexicana A. syringe... There are 2 ml left if you want to try :lol:


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Re: Spore syringe and agar. [Re: enlightenment]
    #24023342 - 01/19/17 04:09 PM (7 years, 11 days ago)

Sometimes a syringe can be too fucked. But rarely is a vendor one not able to be saved by correct plating procedure. You can get away doing the drop on agar with most syringes(i do it all the time) but its piss poor technique


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Invisibleamidogen
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24023353 - 01/19/17 04:12 PM (7 years, 11 days ago)

.


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Edited by amidogen (06/21/18 11:23 AM)


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Re: Spore syringe and agar. [Re: enlightenment]
    #24023374 - 01/19/17 04:19 PM (7 years, 11 days ago)

Quote:

enlightenment said:

I make it a little different and rotate just 2 times



Saw that as someone's signature in advanced mycology, thought it was the most space-efficient use of a petri dish. Since we're taking the smallest piece of the leading edge as soon as we know what we're looking at, I don't see how producing less lab waste is a downside.

Probably 2 rotations are good like enlightenment said because we're not doing single-celled organisms like bodhisatta said. You could also write your name on the plate, I suppose.


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Re: Spore syringe and agar. [Re: AndyHinton]
    #24023429 - 01/19/17 04:41 PM (7 years, 11 days ago)

I like to have a sterilized shot glass. Flame needle squirt out a few drops to cool. A few more into the shot glass. Dip flamed loop into agar the dip into shot glass then back to the agar and make a few Z's


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Offlinefoodguy
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24028487 - 01/21/17 03:38 PM (7 years, 9 days ago)

I did a few drops straight to agar about 5 days ago, no streaking just roll around and it looks like a good bit of bacteria growing as well. I'm about to try the shot glass and streaking in just a few min. I've got plenty of MS left to play with.


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Offlinesynthetik
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24028522 - 01/21/17 03:55 PM (7 years, 9 days ago)

nice... love that shot glass idea, do you wrap in foil or anything?


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Re: Spore syringe and agar. [Re: synthetik]
    #24028665 - 01/21/17 04:59 PM (7 years, 9 days ago)

Yea put them in with any sterilization cycle you do or do them in the oven (dry heat sterilization on Wikipedia)


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Offlinesynthetik
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Re: Spore syringe and agar. [Re: bodhisatta]
    #24038276 - 01/25/17 10:24 AM (7 years, 5 days ago)

nice.. im always finding extra shit to sterilize... really glad I went and just bought a big AA sterilizer to start with from ebay, thing is legit! :smile:


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