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 Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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 Germination help Mexicana & Tampanensis 
#15971533 - 03/20/12 12:00 PM (12 years, 29 days ago) |
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Hoping that someone can offer help or suggestions. 11 days ago I inoculated 10 petri dishes of antibiotic malt extract agar. 2 B+ from syringe, 4 Ps. Mexicana and 4 Ps. Tampanensis both from foil prints. At this time both B+ are close to fully colonized. There is zero apparent grow on either of the Mexicana or Tampanensis dishes. Zero, not a single dish! I believe the prints are a bit over a year old. I plan to just let them sit and see if anything happens. I am wondering if this is normal? Temp is a constant 78 - 80. Is there anything else I can do? I am thinking of creating a syringe with part of the prints and letting the spores hydrate a bit and then going to agar, would this change anything? Should I just have patience?
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#15972081 - 03/20/12 02:35 PM (12 years, 29 days ago) |
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Anyone?
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 TranscendingLife
I Don't Need a Life to Live
 
Registered: 06/09/10
Posts: 21,627
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#15972128 - 03/20/12 02:52 PM (12 years, 29 days ago) |
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Patience, young one. Just like waiting for an answer.
Don't bump your own thread until 24 hours have passed.
&  hydrate the spores before putting them on agar.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
   How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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SomeGuy
I feel better now :)
Registered: 04/18/10
Posts: 7,496
Loc: 
Last seen: 9 years, 10 months
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I'm having the same problem with my P. tampnensis. It's been on MEA for 12 days with no growth and also on whole grain rice for 10 days and not a thing  the print was dated 11/11, so they were pretty fresh, and I left them to hydrate 24 hours before innoculating. I can even see spores in the syringe, but no growth  
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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Re: Germination help Mexicana & Tampanensis [Re: SomeGuy]
#15972211 - 03/20/12 03:09 PM (12 years, 29 days ago) |
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I make sure to let spores that are over a month old rehydrate for 7 days.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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I know I lack patience! This hobby challenges me to work with it. I think I will let the plates sit and in the meantime create a syringe and let the spores hydrate and then try that. Thanks for your help!
Owsley
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mike-corrhizae
Nerd
Registered: 12/21/09
Posts: 131
Loc: Canada 
Last seen: 11 years, 4 months
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#15972532 - 03/20/12 04:15 PM (12 years, 29 days ago) |
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I work with prints only.
try mixing your agar a little wetter next time.
Also cooling your inoculation loupe in the middle of the plate where you intend to streak the spores makes a nice wet divot for them to germinate. This also leaves a little bit of moisture on the loupe when you are scraping from the print making it much easier.
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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Re: Germination help Mexicana & Tampanensis [Re: mike-corrhizae]
#15972924 - 03/20/12 06:01 PM (12 years, 29 days ago) |
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Is there anything that could be done to get the current plates going? Would adding a drop or two of sterilized water help hydrate the spores? I inoculated using a inoculation loop flamed and then cooled in the agar as described above. The agar may have been a bit dryer/stiffer as some boiled over when PCing.
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Persephonesquest
Registered: 11/16/11
Posts: 66
Loc: PNW
Last seen: 11 years, 11 months
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#15976220 - 03/21/12 12:20 PM (12 years, 28 days ago) |
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I am having the same problem with Mexicana. The spores I have are about a year and half old. I created a syring from print and let them hydrate 48 hrs befor putting on agar. It has been 3 weeks and no growth! Really bummed and not sure what to try next?
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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Let them hydrate for 1 week.
& it's time to toss the agar dish & start over.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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I have lift off on one dish! Checked this morning and I can see a tiny white mound of beautiful mycelium in the middle of one of my Tampanensis dishes! I am hoping that this means there is hope for the others or at least one of the Mex! Yea!
Edited by Owsley (03/21/12 12:47 PM)
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Persephonesquest
Registered: 11/16/11
Posts: 66
Loc: PNW
Last seen: 11 years, 11 months
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#15976360 - 03/21/12 12:50 PM (12 years, 28 days ago) |
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Excellent! I think I am starting over and praying
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Persephonesquest
Registered: 11/16/11
Posts: 66
Loc: PNW
Last seen: 11 years, 11 months
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TranscendingLife I will take your advice and hydrate some for a week and see if anything happens, but why would I dump the dishes? There's no contamination issues, wouldn't it make sense to just let them sit and see if anything happens? I've got space. Is it unlikely that anything would happen after 3 weeks?
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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After 3 weeks, it's not really worth it.
You can leave it if you want, but I doubt anything is going to happen w/ it.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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Persephonesquest
Registered: 11/16/11
Posts: 66
Loc: PNW
Last seen: 11 years, 11 months
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Alright, thanks for the help man....just bummed to be starting over and worried that they weren't stored properly and thus won't come alive...
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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dude, I've failed so many times at growing out an iso, it's not even funny.
Persistence & Patience is what this hobby teaches us all.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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Alright still can't get the Mexicana to germinate. Tried hydrating for 7+ days then on fairly wet agar, I also tried directly to rye, just for the hell of it. Both have produced nothing. I'm out of ideas where do I go next? Still have a good amount of spore print. Ideas?
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#16050683 - 04/05/12 11:06 PM (12 years, 13 days ago) |
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try an LC. Lot's of hydration there. That's all you really got left.
--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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Owsley
Artisan of Acid
Registered: 03/20/12
Posts: 359
Loc: Pala
Last seen: 11 years, 3 months
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Ugh, thanks man, I'll give it a try. Nothing left to lose! Any recipes that are known to work better for Mexicana?
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TranscendingLife
I Don't Need a Life to Live
Registered: 06/09/10
Posts: 21,627
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Re: Germination help Mexicana & Tampanensis [Re: Owsley]
#16050847 - 04/05/12 11:43 PM (12 years, 13 days ago) |
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--------------------
AMU: We Quickly Answer Questions Here
"One must accept the probability of failure to experience the elation of success." - TranscendingLife
“A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
How I Do EVERYTHING
"Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung
"Anything that can be done chemically can be done by other means."- William S. Burroughs
"You are as dead now as you will ever be" - Seth
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