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Will they germinate? The Old Spores Thread...


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#81 DickMoby

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Posted 08 November 2020 - 08:07 PM

I'll answer here.

 

Glad to hear you finished the hood!

Was definately worth the two days of work to be able to work with more confidence and comfort.

 

Peroxide agar would maybe work on living tissue, those here didn't germ yet and peroxide would kill everything.

The agar incubator is getting full with all the transfers from the frozen cultures and the spore plates + edible plates, I decided to just buy some antibiotic agar powder and cook a small batch to put those spores on.

 

Not sure where the bacteria came from, only those three plates are dirty, they were the last ones of that 18 plate load from that day, I don't remember anymore in which order they've been inoculated but the Cinctulus print wasn't one of the old prints and has the same contam on the plate.

 

The bacteria started growing from the spore center and did spread through the excess water so it definately came from the spore area, as if the scalpel was dirty and I brought it in with that.

Honestly though I feel comfortable doing agar and wipe and flame between and usually don't have any issues with dirty plates.

 

Gonna get the agar in a couple days and start over.

Don't expect to see any germination on those bacterial plates but gonna keep them on the side for now, just in case.

______________

 

Wanted to ask that earlier, what is H3 and HillBilly ?

Never heard of those


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#82 Freaky

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Posted 08 November 2020 - 08:10 PM

 

Kind of cosmic how we started old H3 spores on the same day Mikey!  :meditate:  

 

What is your agar recipe you used for those ketchup cups? I haven't decided yet if I like my old clear potato/beer/karo agar or this dark activated charcoal mea. 

 

 

I'm sorry I missed the question. The agar pictured and what I started with is an MEAYP.

  • 5gr agar
  • 5gr LME
  • 1 tsp Bakers Yeast
  • 1 fingernail full of tetraclycline
  • 1/2 tsp Peptone
  • 500 ml H2O

Next step is hydrating the spore in syringes with a glycerine H2O solution. Maybe add some glycerine as well to the agar.

 

 

 

 

This is great info, thank you for the agar recipe! I forgot yeast on my first round but I won't on the next round I do.  I'll either do some transfers or just toss all of these and start over again.

 

Was surprised to find something growing in the PE and PFA from the old sporeworks syrings.  It's hard to tell but I'm not so sure it's mycelium germination - after this long how likely to get a contam in the agar though? Best only way to tell is to do some transfers and see what comes of it.  

 

PE Agar

PE Agar1 10-25.jpg

PE Agar2 10-25.jpg

 

PFA Agar 

PFA Agar1 10-25.jpg


Edited by Freaky, 08 November 2020 - 08:11 PM.


#83 DickMoby

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Posted 08 November 2020 - 08:15 PM

The first two look like myc to my eye, not sure about the third one tho.

Nice!


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#84 Freaky

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Posted 08 November 2020 - 08:29 PM

I'll answer here.

 

Glad to hear you finished the hood!

Was definately worth the two days of work to be able to work with more confidence and comfort.

 

Peroxide agar would maybe work on living tissue, those here didn't germ yet and peroxide would kill everything.

The agar incubator is getting full with all the transfers from the frozen cultures and the spore plates + edible plates, I decided to just buy some antibiotic agar powder and cook a small batch to put those spores on.

 

Not sure where the bacteria came from, only those three plates are dirty, they were the last ones of that 18 plate load from that day, I don't remember anymore in which order they've been inoculated but the Cinctulus print wasn't one of the old prints and has the same contam on the plate.

 

The bacteria started growing from the spore center and did spread through the excess water so it definately came from the spore area, as if the scalpel was dirty and I brought it in with that.

Honestly though I feel comfortable doing agar and wipe and flame between and usually don't have any issues with dirty plates.

 

Gonna get the agar in a couple days and start over.

Don't expect to see any germination on those bacterial plates but gonna keep them on the side for now, just in case.

______________

 

Wanted to ask that earlier, what is H3 and HillBilly ?

Never heard of those

 

I've never worked with antibiotic agar of any kind so that will be a good learning curve for me what you're able to share using it and the level of success for you.

 

Hillbilly is a strain of cubensis from an old sponsor/vendor here - it was his own strain. I'm not sure what happened to him or if he's still in business or not.

H3 was a strain created in honor of Hippie3 - a cross of his favorite strains - Dixie and Texas? I can't remember which strains created H3......Mikey knows though

 

And yes, I'm feeling confident about the PE agar jars.  I'll do those transfers in a couple days with them.  I want to cook some fresher agar and include the yeast in the next batch.



#85 coorsmikey

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Posted 08 November 2020 - 09:43 PM

Well its was Dixieland and Hillbilly from what I recollect. I have had a couple of yeast fermented drinks so if I'm mistaken I will come back and correct soon. I remember the strains but what I don't recall is how the cross was done. That would be some good reads.


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#86 Freaky

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Posted 08 November 2020 - 09:56 PM

Well its was Dixieland and Hillbilly from what I recollect. I have had a couple of yeast fermented drinks so if I'm mistaken I will come back and correct soon. I remember the strains but what I don't recall is how the cross was done. That would be some good reads.

 

I was on hiatus during the H3 creation so I am not sure but Dixie and Hillbilly strain would make the most sense.  I know Hip loved all three of those strains though, Dixie, Hillbilly and Tex - maybe why I thought Tex. 

 

I may dig around for some H3 grow logs too maybe be able to gather some info or get an idea of how the look.  Dixie's were always one of my favorites - really a pretty strain.



#87 Freaky

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Posted 08 November 2020 - 09:58 PM

https://mycotopia.ne...hippie3-strain/

 

I guess I should of just checked the strain base in the vaults.  The kind Dr. has a great thread! Too bad my memory is going in my old age but I definitely had a post in there and have forgotten the details.

It's Dixie and Texas strains


Edited by Freaky, 08 November 2020 - 10:01 PM.

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#88 Freaky

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Posted 08 November 2020 - 10:09 PM

https://mycotopia.ne...hippie3-strain/

 

MaH - the H3 syringe I am working with is definitely from the old Mycrotopia! This makes me very happy and even more determined to get that syringe germinating.


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#89 cujoloki

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Posted 09 November 2020 - 02:39 AM

Whoa that's gonna be a healthy plate. I would love to try to revive those pink buffalo you haven't thrown them away. From my perusing I'm guessing all the PE are called for. I'm back into the hobby and working on my agar game.That said it would be cool to join you guys in trying to run some old stock.

Edited by cujoloki, 09 November 2020 - 02:40 AM.

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#90 DickMoby

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Posted 09 November 2020 - 09:06 AM

Uuh I like those caps, looks like a nice variety!

 

next question, sry 

what are Hillbilly and Dixie ?

:rolleyes:



#91 Freaky

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Posted 10 November 2020 - 12:04 AM

Uuh I like those caps, looks like a nice variety!

 

next question, sry 

what are Hillbilly and Dixie ?

:rolleyes:

 

We have Dial8  :wub:  to thank for Dixie - https://mycotopia.ne...xieland-strain/  a few posts down in this thread Dial8 explains the origins

 

 

Hillbilly if memory serves is also another wild cube worked with until it was tamed to be a great mushroom strain for invitro growing as well as on bulk. I remember way back when Hillbilly offered up pint jars full of spore solution to members who donated to Mycotopia - I had one of those jars and it noc'd up so much grain lol - Hillbilly (who also named his strain Hillbilly) was a vendor as well for a time - The Good Spore I think his site was.  Not sure if he's still around or still in the spore business.  There are some good Hillbilly grow threads in the Vaults.


Edited by Freaky, 10 November 2020 - 12:09 AM.

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#92 Freaky

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Posted 10 November 2020 - 12:06 AM

Whoa that's gonna be a healthy plate. I would love to try to revive those pink buffalo you haven't thrown them away. From my perusing I'm guessing all the PE are called for. I'm back into the hobby and working on my agar game.That said it would be cool to join you guys in trying to run some old stock.

 

Not sure if I still have the PB - but I've still got a lot of old prints not claimed.  I pm you.


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#93 Freaky

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Posted 11 November 2020 - 12:44 AM

Did some transfers tonight of the PE and PFA.  There are still other jars starting to germinate so I'll keep working with them and hopefully get something worthwhile.

 

PE PE 2010 sporeworks MSS.jpg

PFA PFA - 2010 Sporeworks MSS.jpg

 

12 total transfers completed PE & PFA Transfers(1) 11-10.jpg

 

 

I think I may start over on the H3 - the bacterial contam is pretty fierce in them so I'm going to try again.  I'll get started on some of the prints also now the hood is good.


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#94 DickMoby

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Posted 12 November 2020 - 06:10 PM

Well I kept the bacterial plates and possibly there is some visible germination

 

IMG_20201112_212440.jpg

 

it's really hard to see and also could be the bacteria showing up from underneath the spores, couldn't see it exactly, even with a really bright LED and magnifying glass its hard to get a sharp view.

 

Pan Cinctulus similar, can't say exactly if the grey spots are germination or just the white bacteria showing up from underneath.

 

Goliath does nothing at all.

 

Just cooked a batch of antibiotic agar and gonna scrape a bit of those sectors surfaces to the medicinal agar and check if some myc grows from it.

 

Also gonna start all three PE, pan Cinc and Goliath again directly on the antibiotic agar.


Edited by DickMoby, 12 November 2020 - 06:14 PM.


#95 Freaky

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Posted 13 November 2020 - 12:41 AM

My H3 plates - the bacterial contams took over - there is still one that looks like I may be able to grab a small piece of healthy myc.  Hopefully that antibiotic agar does the trick on some of those prints.  

 

My PE and PFA transfers I'll be checking on tomorrow - another of the original PE and PFA's have germination so I'll do another set of transfers from those also.  Hoping this weekend to do a lot of prints to agar and see how it grows.



#96 DrepsiLocybe

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Posted 13 November 2020 - 12:55 PM

My H3 plates - the bacterial contams took over - there is still one that looks like I may be able to grab a small piece of healthy myc. Hopefully that antibiotic agar does the trick on some of those prints.

My PE and PFA transfers I'll be checking on tomorrow - another of the original PE and PFA's have germination so I'll do another set of transfers from those also. Hoping this weekend to do a lot of prints to agar and see how it grows.

im struggling with the yosterizii and pesa.
i recently got a hold of gentamicin powder and made few plates with it but im not 100% sure on the ratio...
so far im trying 500ml water/ 10g agar/5g lme/ 0.2g gentamicin. (is the antibiotic ratio right? first time working with it)
also i put the spore in centrifugal tubes to hydrate for a week or so. ill be trying to inoculate some plates with them again this weekend.
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#97 Freaky

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Posted 13 November 2020 - 01:14 PM

im struggling with the yosterizii and pesa.
i recently got a hold of gentamicin powder and made few plates with it but im not 100% sure on the ratio...
so far im trying 500ml water/ 10g agar/5g lme/ 0.2g gentamicin. (is the antibiotic ratio right? first time working with it)
also i put the spore in centrifugal tubes to hydrate for a week or so. ill be trying to inoculate some plates with them again this weekend.

 

 

 

 Thanks for sharing an update DrepsiLocybe! I've never used the antibiotic agar or gentamicin - hopefully someone can give us some info on that. I'll look around though to see if there are old archives using it.

 

I'll be doing a lot of agar work this weekend and hopefully can get some spores to germinate.  I've been adding the yeast and charcoal powder to my MEA and the 10 year old spore syringes seem to really be doing okay with that.  If I can get clean transfers away from bacteria and other baddies hopefully I can pull off something decent without going the antibiotic route but it's doubtful right now.  

 

I'll be updating and checking on the old Acadia Coast print I dropped on agar and see if anything besides contams are germinating on it.


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#98 DickMoby

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Posted 16 November 2020 - 10:17 AM

Found that

 

http://cshprotocols....?text_only=true

____________

 

Not gonna use those crappy big plates from amazon anymore, even though they're made of boro, the quality is so bad that it's not worth working with them.

 

Started the antibiotic agar 3 days ago, the premix contains: Potassium dihydrogen phosphate, chloramphenicol, peptone, yeast extract and malt.

 

IMG_20201115_173403.jpg

 

For some reason condensation pooled right on the spores which wasn't on purpose because those were inoculated with plain spores, not with a syringe but honestly I think that might be a good idea to do - put spores on the plate and drip a drop of sterilized water on top instead of the prehydration.

 

PE aswell as some others were also put on fresh plates, gonna check the plates every day 


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#99 Freaky

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Posted 16 November 2020 - 12:30 PM

I like that idea too - drop sterile water on the spores - will be nice if those germinate. 

 

I was busy working and spending family time this weekend so tonight I'll be updating as well as putting some old prints to agar. 


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#100 Freaky

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Posted 27 November 2020 - 01:26 AM

Not anything exciting to update with but I'm moving along with the spore syringes from 2010.  I've done 2nd and 3rd transfers on the PE and PFA both. Today I had some extra popcorn jars that I decided to use with some of the agar plates (jars) rather than use with clone tissue. 

 

The PE was a 2nd transfer plate (jar) that seemed to be moving along nicely and didn't show any contams so why not see what it does on grain.

PE 2C (1st transfer) 11-8-2020.JPG

PE 2C to pcorn 11-27-2020.JPG

 

The PF Albino plate (jar) is a from the ms syringe - it took it a while to get going but it looked great so it went into 2 half pint jars of popcorn. There are some 2nd and 3rd transfers (but not from this plate) I will be working with also.  

PF Albino (2010 ms syringe)10-25-2020.JPG

 

 

As for the prints - no luck yet, but I'll be trying some new agar this weekend and hopefully get something to germinate.  It may be a lost cause on prints up to 15 years old. 


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