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Rosen_Rot
Learning



Registered: 12/06/14
Posts: 1,225
Loc: Goa
Last seen: 11 months, 22 days
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Agar taking too long to germinate spore?
#23837820 - 11/16/16 06:32 AM (7 years, 2 months ago) |
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10/11/16 I inoculated 13 petri dishes with CFA, 10g cat food, 10g agar agar, 500ml water. Pced for 30mins after the rocker shakes, cooled down and then poured in front of a laminar starting from the bottom up
I followed standard procedure, burning the syringe needle till it's red hot on an alcohol lamp, pushing the plunger to cool down the needle and pushed the plunger again to drop 3 drops of B+ MS onto the centre of my agar mix
Its been 6 days and I haven't seen any mycelium development. There doesn't seem to be any mold growing on the agar either. Quite possibly some agar plates might have bacteria as I see on some sections of petris with a small uprised agar bubble.
They are marked with a red circle.
Some of the petris have like powder on the outside of the petri lid but that is okay cause it's just from my powdered surgical gloves. Some of my petris also have condensation so it might look milky. I assure you on the inside they look like it's just agar.
The syringe was a bit old, I bought it last year and stored it in the fridge where it stayed at 4C all summer
How long does it usually take on agar for spores to germinate??
Did I over load the agar dishes with spores and is that why they won't germinate??
Should I throw them away?? 
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"The internet has one rule; use or be used" - Bjeldiablo ''there is no loneliness, only moments where contentment is fleeting'' SBJs "The Basics" 3iRiS9 "Cirque du freak" B+ BONANZA
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enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 22 days
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Re: Agar taking too long to germinate spore? [Re: Rosen_Rot]
#23837825 - 11/16/16 06:38 AM (7 years, 2 months ago) |
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Wait a few more days. Old spores can take more time to germinate IME. I had massive problems with old spore prints. I made syringes out of the old prints and it helped me a lot (hydration). But you already have a syringe. You can try to inoculate BRF paste.
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Rosen_Rot
Learning



Registered: 12/06/14
Posts: 1,225
Loc: Goa
Last seen: 11 months, 22 days
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Re: Agar taking too long to germinate spore? [Re: enlightenment]
#23837831 - 11/16/16 06:41 AM (7 years, 2 months ago) |
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I inoculated more petris this morning with a KSSS swab and also waiting for some agar to finish curing before I make more plates with whatever is left in the B+ syringe.
Luckily I got some grain jars so if all else fails I can always introduce a colonized grain.
It just sucks when wasting so much dishes and time
BTW
Agar plates are being stored at 21-22C
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"The internet has one rule; use or be used" - Bjeldiablo ''there is no loneliness, only moments where contentment is fleeting'' SBJs "The Basics" 3iRiS9 "Cirque du freak" B+ BONANZA
Edited by Rosen_Rot (11/16/16 06:41 AM)
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enlightenment
alchemist


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Re: Agar taking too long to germinate spore? [Re: enlightenment]
#23837842 - 11/16/16 06:51 AM (7 years, 2 months ago) |
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That would have been my next question... temperature is fine. I know the pain.. Tossed 20 dishes after trying to get growth from a Psi. mexicana A print.
Some of the dishes look like they show signs of growth IMO. It often starts to look milky where you inoculated. Can you make single pictures?
Especially the dishes you marked with red circles look like they started to germinate.
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Mycolorado
Hobbyist


Registered: 07/23/16
Posts: 8,529
Loc: Interdimensional Bootcamp
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Re: Agar taking too long to germinate spore? [Re: enlightenment]
#23837857 - 11/16/16 07:05 AM (7 years, 2 months ago) |
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Why CFA? Try a different agar recipe. This is the first I've seen anyone using CFA. FH mentions it but I don't think it was his standard, more of an experiment it seemed. Either way, you might try switching to something a little more in line with what is typically used...MEA, DFA, PDYA, etc. cat food is super high in protein and fat.
Also, cubes typically germinate in 7 - 10 days.
Edited by Mycolorado (11/16/16 07:11 AM)
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Rosen_Rot
Learning



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Re: Agar taking too long to germinate spore? [Re: Mycolorado]
#23838346 - 11/16/16 10:28 AM (7 years, 2 months ago) |
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From my research, I believe it was a post by Pasty and a bunch of other agar recipe threads, CFA had some very interesting results and proved to be a very viable cheap source of media to cultivate mycelium. For me it was the easiest to acquire to my circumstance, my mother keeps cats so we get high quality cat food like WHISKAS..etc
I tried looking for LME and ME but couldn't find a good decently priced source. All online sources proved to be very expensive or just not the right stuff I was looking for.
I will be trying DFA, any recommendation as to brand?
Enlightenment; I wasn't sure if the milky streaks within the plates were mycelium development, condensation or mold from taking too long to germinate. Glad to know that they might be showing signs of life, that's a step in the right direction 
I will try and get individual pictures tomorrow evening so look out for this thread
And thank you to anyone responding to this thread, I appreciate the help
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"The internet has one rule; use or be used" - Bjeldiablo ''there is no loneliness, only moments where contentment is fleeting'' SBJs "The Basics" 3iRiS9 "Cirque du freak" B+ BONANZA
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: Agar taking too long to germinate spore? [Re: Mycolorado]
#23838408 - 11/16/16 10:51 AM (7 years, 2 months ago) |
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Quote:
Mycolorado said: Why CFA? Try a different agar recipe. This is the first I've seen anyone using CFA. FH mentions it but I don't think it was his standard, more of an experiment it seemed. Either way, you might try switching to something a little more in line with what is typically used...MEA, DFA, PDYA, etc. cat food is super high in protein and fat.
Also, cubes typically germinate in 7 - 10 days.
Do a search on cat and dog food agar. They were the rage here a year or 2 ago
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Mycolorado
Hobbyist


Registered: 07/23/16
Posts: 8,529
Loc: Interdimensional Bootcamp
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Re: Agar taking too long to germinate spore? [Re: Mad Season]
#23838428 - 11/16/16 10:59 AM (7 years, 2 months ago) |
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I use DFA all the time and suggested the OP try it. What's that got to do with CFA being all the rage a couple years ago. If it were good, it would still be the rage today. But thanks for the useful tip.
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Panarchist
Cyanescendant



Registered: 04/02/16
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Loc: Copeland
Last seen: 7 months, 17 days
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Re: Agar taking too long to germinate spore? [Re: Rosen_Rot]
#23840046 - 11/16/16 08:56 PM (7 years, 2 months ago) |
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Quote:
Rosen_Rot said:
I tried looking for LME and ME but couldn't find a good decently priced source. All online sources proved to be very expensive or just not the right stuff I was looking for.

i got some Seaweed Solutions brand MEA off Ebay recently. it wasn't much more than the food grade agar. the food grade would have been $12.00 for 100 grams. the MEA was $16.95.
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blindingleaf
blue collar underworld


Registered: 07/19/13
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Loc: sub-surface unseen
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Re: Agar taking too long to germinate spore? [Re: Panarchist] 1
#23840773 - 11/17/16 05:52 AM (7 years, 2 months ago) |
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Quote:
my mother keeps cats so we get high quality cat food like WHISKAS
DONT PASS ME NO REGULAR!!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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The Mycologist
Explorer

Registered: 05/06/16
Posts: 3,024
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Re: Agar taking too long to germinate spore? [Re: blindingleaf]
#23840799 - 11/17/16 06:21 AM (7 years, 2 months ago) |
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You only need one drop of spores solution, there are so many spores in even a clear section of the sryinge. They should show growth soon.
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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ComebackKid
Multispore Enthusiast



Registered: 05/27/16
Posts: 3,951
Loc: ked in the trunk of a car
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Re: Agar taking too long to germinate spore? [Re: The Mycologist]
#23840831 - 11/17/16 06:52 AM (7 years, 2 months ago) |
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I feel your pain. Been trying to get spores to germinate on agar for a couple months now  I just recently put them in a syringe and dropped on both agar and BRF paste which I hear is a good way to germ before moving to agar.
My AA+ spores on the other hand took ~2 weeks to germ. Just be patient for now, I wouldn't rule them out as duds just yet!
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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Rosen_Rot
Learning



Registered: 12/06/14
Posts: 1,225
Loc: Goa
Last seen: 11 months, 22 days
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Re: Agar taking too long to germinate spore? [Re: ComebackKid]
#23841869 - 11/17/16 01:58 PM (7 years, 2 months ago) |
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Hey guys sorry for the poor quality but I was in a hurry. Looks like I am starting to see whisps of mycelium starting to form and I think they are well and alive 
Quote:
ComebackKid said: I feel your pain. Been trying to get spores to germinate on agar for a couple months now  I just recently put them in a syringe and dropped on both agar and BRF paste which I hear is a good way to germ before moving to agar.
My AA+ spores on the other hand took ~2 weeks to germ. Just be patient for now, I wouldn't rule them out as duds just yet!
I was under the impression that on agar spores would germinate within a few days I guess I'm still impatient but I think I am yielding some good results
Here are some pics

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"The internet has one rule; use or be used" - Bjeldiablo ''there is no loneliness, only moments where contentment is fleeting'' SBJs "The Basics" 3iRiS9 "Cirque du freak" B+ BONANZA
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ComebackKid
Multispore Enthusiast



Registered: 05/27/16
Posts: 3,951
Loc: ked in the trunk of a car
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Re: Agar taking too long to germinate spore? [Re: Rosen_Rot]
#23842874 - 11/17/16 06:07 PM (7 years, 2 months ago) |
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Congrats man! Yeah, I've read that spores can take up to 18 days on agar sometimes. If this hobby has taught me anything, it's a lesson in patients
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Substrate surface conditions / Monotub prep and care
Look around you... Everything you see exists inside the mind. Consciousness, the awareness that is experiencing this mind, is peering in from outside the universe. Our individual experiences are all part of the universe's experience of itself
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enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
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Re: Agar taking too long to germinate spore? [Re: ComebackKid]
#23843904 - 11/18/16 12:26 AM (7 years, 2 months ago) |
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 Good that you did not toss them. They already germinated on your first post. Those milky spots i wrote about. Good luck with your dishes and further projects
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Rosen_Rot
Learning



Registered: 12/06/14
Posts: 1,225
Loc: Goa
Last seen: 11 months, 22 days
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Re: Agar taking too long to germinate spore? [Re: enlightenment]
#23843936 - 11/18/16 12:53 AM (7 years, 2 months ago) |
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Super happy thank you so much for your input fellas really appreciate it!!
Unfortunately I wasn't so fortunate with the green elf cup fungus, only one plate left and seems inactive for now
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"The internet has one rule; use or be used" - Bjeldiablo ''there is no loneliness, only moments where contentment is fleeting'' SBJs "The Basics" 3iRiS9 "Cirque du freak" B+ BONANZA
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