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InvisibleGorlami
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Ps. Galindoi Spore germination on agar
    #19347940 - 12/30/13 06:58 PM (10 years, 1 month ago)

Hello friends,

I have a print of ATL7 that i've scraped over agar on Dec 17th and so far, nothing.

Does this particular strain take longer to germinate compared to Cubensis?  I've inoculated a plate on the same date with a MS syringe of GT's and it took a matter of days for something to occur, so I know I can't be messing up the recipe.  I'm using potato infused water with unpasteurized honey as my agar recipe.  Might be too nutritious, I see thick cottony growth without much rhizomorphic growth happening, would this matter for a different strain such as Galindoi?

Thanks in advance.  :cheers:


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OfflinecronicrFacebook
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19347958 - 12/30/13 07:01 PM (10 years, 1 month ago)

did u use a loop or a blade?


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: cronicr]
    #19347970 - 12/30/13 07:03 PM (10 years, 1 month ago)

I used an agar cutting tool, looks like a spear.  Both sides are not sharpened.


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OfflinecronicrFacebook
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19347979 - 12/30/13 07:05 PM (10 years, 1 month ago)

as long as it isn't contaminating all you can really do is wait it out, been awhile since i did any scletoria but as far as i remember my germination was about 2 weeks


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: cronicr]
    #19348007 - 12/30/13 07:09 PM (10 years, 1 month ago)

No contaminations as of yet.

The prints were tiny, I scraped the spores off sparingly, then wiped the tool on the agar.

What is the best way to proceed in order to get the best results when transfering spores from a print to agar?

:bigblunt:


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OfflinecronicrFacebook
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19348021 - 12/30/13 07:13 PM (10 years, 1 month ago)

i like to dip my loop in the agar first to cool it, also helps to get spores stuck to the loop:thumbup:


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: cronicr]
    #19348060 - 12/30/13 07:20 PM (10 years, 1 month ago)

Hmmm, so after flaming the innoc loop, you dip in the agar first to cool it and somewhat moisten it?  Then touch your print, then back to the agar?

I'm simply taking my print on aluminium, and scraping the print with my slicing tool over the agar.  :S

I'm using 1/2 pints as agar vessels, so they have quite a way to fall.  Ugh I hope this worked... not akin to working with prints. :frown:

EDIT:  Doing this in a SAB


Edited by Gorlami (12/30/13 07:21 PM)


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OfflinecronicrFacebook
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19348073 - 12/30/13 07:21 PM (10 years, 1 month ago)

Quote:

whalechunks said:
Hmmm, so after flaming the innoc loop, you dip in the agar first to cool it and somewhat moisten it?  Then touch your print, then back to the agar?




:thumbup:


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: cronicr]
    #19391439 - 01/09/14 12:27 AM (10 years, 22 days ago)

It has now been three weeks without germination on agar.  When can I safely say they won't ever germinate?  There aren't any contamns as of yet, so I at least have that going for me.  I've hear of germination taking up to 3 weeks, any chance it could take even longer?! 

I scrapped my spores with an agar scalpel over a 1/2 pint jar that had the agar media.

Agar consisted of 10gr Agar Agar, 500ml with potato water (boiled 30 minutes), with teaspoon of unpasteurized honey.

Had no problem inocculating with 1 drop off another syringe, to which the mycelium showed up within 3 days.

The wait is killing me!!!!  After reading all the rave comments and reviews on their effects, I can't wait to try them for myself. :trippnballs:


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Offlinegornyhuy
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19392537 - 01/09/14 07:39 AM (10 years, 22 days ago)

How old was the print?

I have used the same agar recipe and scraping technique plenty of times with cubes and had very fast growth, just like you've seen.  I was planning on doing the same with sclerotia spores this weekend, but now you've got me worried.

Please update this thread with any progress or solutions, I'll be watching!

Thanks...


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InvisibleSkinty
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Re: Ps. Galindoi Spore germination on agar [Re: cronicr]
    #19392548 - 01/09/14 07:43 AM (10 years, 22 days ago)

Quote:

cronicr said:
did u use a loop or a blade?




Hey cronicr why do you ask that? Is there a reason not to use a blade? I've been using my scalpel - red hot, cool in agar, scrape some spore and place on agar - and been getting mixed results (with spores)... :confused::thumbup:


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Offlinealabamamushie
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Re: Ps. Galindoi Spore germination on agar [Re: Skinty]
    #19392558 - 01/09/14 07:45 AM (10 years, 22 days ago)

Quote:

you can swipe or acrape thats really up to u it just controls how much spores youuse, i have lots so i don't mind scraping and dumping a bunch in there lol,as for ge as long as it has enough not to seal and create a vacuum it should be fine

had a conversation with cron about this yesterday :laugh:


@ gorny dont be scared of failure failure is progress towards Success IMO


Edited by alabamamushie (01/09/14 07:47 AM)


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Offlinegornyhuy
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Re: Ps. Galindoi Spore germination on agar [Re: alabamamushie]
    #19392825 - 01/09/14 09:39 AM (10 years, 22 days ago)

Quote:

alabamamushie said:
Quote:

you can swipe or acrape thats really up to u it just controls how much spores youuse, i have lots so i don't mind scraping and dumping a bunch in there lol,as for ge as long as it has enough not to seal and create a vacuum it should be fine

had a conversation with cron about this yesterday :laugh:


@ gorny dont be scared of failure failure is progress towards Success IMO




I've had more success by doing a quick light scalpel scraping of prints on foil, swiping with a loop often doesn't germinate for me... maybe my spores are really stuck on there.

@Alabam - thanks! I'm not scared, but I hate wasting effort and supplies and most of all TIME with a technique that may not work well for sclerotia for whatever reason.

Seems to me spores are spores, and I know plenty will get on the dish.  So my question is - does the potato/honey agar not work as well for sclerotia for some reason?


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InvisibleinskiM
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Re: Ps. Galindoi Spore germination on agar [Re: gornyhuy]
    #19393014 - 01/09/14 10:25 AM (10 years, 22 days ago)

If you still have spores to work with try again but don't use honey, it has anti fungal properties.
Make your agar recipe as before but use dextrose instead of honey.


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Offlineblueconfusion
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Re: Ps. Galindoi Spore germination on agar [Re: inski]
    #19393053 - 01/09/14 10:39 AM (10 years, 22 days ago)

I have some ALTs on PDYA recipe linked in my sig they love it and are moving quickly


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OfflinecronicrFacebook
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Re: Ps. Galindoi Spore germination on agar [Re: alabamamushie]
    #19393055 - 01/09/14 10:39 AM (10 years, 22 days ago)

Quote:

inski said:
If you still have spores to work with try again but don't use honey, it has anti fungal properties.
Make your agar recipe as before but use dextrose instead of honey.



:thumbup:
Quote:

Skinty said:
Quote:

cronicr said:
did u use a loop or a blade?




Hey cronicr why do you ask that? Is there a reason not to use a blade? I've been using my scalpel - red hot, cool in agar, scrape some spore and place on agar - and been getting mixed results (with spores)... :confused::thumbup:



scalpels are sharp and damage spores easy, i use something dull to scrape
Quote:

alabamamushie said:
Quote:

you can swipe or acrape thats really up to u it just controls how much spores youuse, i have lots so i don't mind scraping and dumping a bunch in there lol,as for ge as long as it has enough not to seal and create a vacuum it should be fine

had a conversation with cron about this yesterday :laugh:


@ gorny dont be scared of failure failure is progress towards Success IMO



:rockon:


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It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: inski]
    #19394143 - 01/09/14 03:13 PM (10 years, 21 days ago)

Quote:

gornyhuy said:
How old was the print?

I have used the same agar recipe and scraping technique plenty of times with cubes and had very fast growth, just like you've seen.  I was planning on doing the same with sclerotia spores this weekend, but now you've got me worried.

Please update this thread with any progress or solutions, I'll be watching!

Thanks...




Hey Gornyhuy!  Fancy seeing you here! :smile:

To answer your question, I have no idea how long they've been sitting at the vendors... but it took me 1 month before using them.  Considering the high demand of these particular prints, i'm assuming they didn't linger long before being shipped.

However, it's reassuring to know you've had success with this technique. :smile:  Hopefully this is just a test of my patience.




Quote:

inski said:
If you still have spores to work with try again but don't use honey, it has anti fungal properties.
Make your agar recipe as before but use dextrose instead of honey.




But i've had spores germinate on this recipe with a spore syringe within 3 days.  I'm assuming it's quicker because the spores are already hydrated?

I'm almost tempted to squirt some sterile water on my agar plate to hydrate those suckers... but i'm sure folks here will caution me not to. :tongue:


Thanks everyone for submitting their opinions and concerns.


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: blueconfusion]
    #19394152 - 01/09/14 03:16 PM (10 years, 21 days ago)

Quote:

blueconfusion said:
I have some ALTs on PDYA recipe linked in my sig they love it and are moving quickly





Hey blue, you're tek was actually the blueprint for my agar recipe, I just didn't have any dextrose or yeast hanging around.  If i'm not mistaken your TEK even mentions using honey as a dextrose substitute? :laugh:


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Offlineblueconfusion
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19395305 - 01/09/14 06:50 PM (10 years, 21 days ago)

It does but remember honey is a preservative, I have never used it myself in agar, but I have seen it used in other recipes.  I've used it in LCs but like I said it is a preservative so use sparingly.


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InvisibleGorlami
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Re: Ps. Galindoi Spore germination on agar [Re: blueconfusion]
    #19445986 - 01/20/14 04:21 AM (10 years, 11 days ago)

Quote:

blueconfusion said:
It does but remember honey is a preservative, I have never used it myself in agar, but I have seen it used in other recipes.  I've used it in LCs but like I said it is a preservative so use sparingly.





Thanks, that may have been the problem in the end.  That or crappy technique.

I made another agar batch but with Dextrose this time, spores germinated in 2 days. Wowzers! :laugh:

Thanks to everyone who chimed in, couldn't ask for more! :smile:


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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19446154 - 01/20/14 05:49 AM (10 years, 11 days ago)

I had the same issue. While making spawn for another project I just threw in a couple hardly filled 1/2 pint jars of rye berries and scrapped some spores directly onto them. For what ever reason they had no issues germinating onto that... you'd think that it would be thriving on the agar. Been missing with different amount of agar/nutrients now so maybe that will solve the issue. Anyway, I let that grow out a bit then simply just scooped a grain up and threw that on agar...

Id still like to get it to germinate on agar tho, thinkin I may try some PDA or check around and see what others are having success on. MEA has worked well for me but its always spores that give me a hit or miss on germination. Best of luck!


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Offlinegornyhuy
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Re: Ps. Galindoi Spore germination on agar [Re: Gorlami]
    #19446699 - 01/20/14 09:01 AM (10 years, 11 days ago)

Quote:

whalechunks said:
Quote:

blueconfusion said:
It does but remember honey is a preservative, I have never used it myself in agar, but I have seen it used in other recipes.  I've used it in LCs but like I said it is a preservative so use sparingly.





Thanks, that may have been the problem in the end.  That or crappy technique.

I made another agar batch but with Dextrose this time, spores germinated in 2 days. Wowzers! :laugh:

Thanks to everyone who chimed in, couldn't ask for more! :smile:




Thanks for the update!  I have just gotten a big bag of LME and am about to make my first dextrose based agar and probably LC while I'm at it.  Very interested to see how it performs versus the old potato + honey recipe.


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