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Crossing strain varieties via swab to spore solution


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#281 rockyfungus

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Posted 03 April 2021 - 01:12 PM

My little experiment doesn't tell me anything so far. Interesting to see some variation in myc. I tried taking photos but my DIY agar plates are impossible to share.

 

Let me try and explain this again. I have no clue if I'm even replicating Faht's. I made swabs from some AA+ and PE grown very recently. They are not swabbed together per Faht. 

 

I have 2 "control plates" AA+ and PE. AA+ appears more aggressive and has more cottony growth. PE forms a central spoke. 

So 1 plate was 1 swab of each rubbed together in an X (the central point hopefully being best candidate)

Another plate is the individual swabs not rubbed together in the same X.

All of the used tips were then rubbed together in sterile water and a syringe was repeatedly filled/emptied in just enough solution to make 2 other test plates. Test places were inoculated with 1 drop and swirled...(too lazy for loop)
Every plate that is "mixed" has both sets of growth and I'm going to try and isolate from the central X.

I'll transfer out the central overlap of the X or any growth that appears much different from the control plates. Add grain to the T0's. If the T1's are clean go for the same. Hoping to fruit right from my agar plates. I'll have to do some traditional jars to be safe...


Edited by rockyfungus, 03 April 2021 - 01:13 PM.

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#282 fahtster

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Posted 03 April 2021 - 02:09 PM

For sure. I think I'm gonna make four little take out trays and try a mixed casing of it and coir and then two others of just orchid moss. Two with a 1/4" casing and two with something more like 1/2".
The only real upside I'm hoping to find is better aeration and no need to add any garden lime or the such.
Have you ever used diamateous earth? Or however that is spelled?
Even if it ain't all good. It's all good enough.

I’ve heard of ppl using that to combat bug infestation but not much beyond that.. again, doesn’t mean it won’t work.. I’m sure there’s at least a few untapped potential materials out there that would be very effective for casings. I like the project.

My little experiment doesn't tell me anything so far. Interesting to see some variation in myc. I tried taking photos but my DIY agar plates are impossible to share.
 
Let me try and explain this again. I have no clue if I'm even replicating Faht's. I made swabs from some AA+ and PE grown very recently. They are not swabbed together per Faht. 
 
I have 2 "control plates" AA+ and PE. AA+ appears more aggressive and has more cottony growth. PE forms a central spoke. 
So 1 plate was 1 swab of each rubbed together in an X (the central point hopefully being best candidate)
Another plate is the individual swabs not rubbed together in the same X.
All of the used tips were then rubbed together in sterile water and a syringe was repeatedly filled/emptied in just enough solution to make 2 other test plates. Test places were inoculated with 1 drop and swirled...(too lazy for loop)
Every plate that is "mixed" has both sets of growth and I'm going to try and isolate from the central X.
I'll transfer out the central overlap of the X or any growth that appears much different from the control plates. Add grain to the T0's. If the T1's are clean go for the same. Hoping to fruit right from my agar plates. I'll have to do some traditional jars to be safe...


I like your project as well.. lots of controls which is nice. I’d definitely take from where the streaks overlap and grow that out.. not sure how much the appearance of myc growth is going to tell you.. especially with ms.. could steer you in the right direction but I’ve seen all types of growth even within a single variety. I’ve had tometose (sp?) myc do great and rhizo myc do poorly.. the apeu I have is extremely tom and puts out banging flushes. Anyway.. a lot of ppl do long snake like streaks of one variety and then take the other swab and streak straight down through the zig zags a few times to get as many overlaps as possible.. just something to think about for later.

I wouldn’t worry too much about the swabs not being layered.. I think that rubbing them together at the end to make spore solution is going to be nice and effective for making new spore clumps.. maybe not as much so as if they had the varieties layered on top of each other, but still much better than just streaking over one another on a plate. Nice job and carry on!

Faht

Edited by fahtster, 03 April 2021 - 02:23 PM.

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#283 DarkNchildlike

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Posted 03 April 2021 - 03:00 PM

@rockyfungus , that's so awesome that you are trying to do some replications and variations on the method I can't stress that enough.

I gotta get some swabs and I'll try taking swabs on the one side 2 at a time side by side and letting dry in original package for a week then using same swabs to take on new spores from different strain and repeat drying. And will plan further from there.

Right now I am on second flush of four trays in my Martha what I believe is oakridge, but I am not positive. Had a mix up. Beyond that I have a ton of straw that is colonizing of South American variety. I just did a scraping of spores from AA+, Fahtsters PfRedspore, (the MS canopy he pulled, in his pic where print he sent came from looked awesome had to do it lol) LizardKing from Baphomet, and Alacabenzi (so may people have said it's their favorite in the past so I wanted to give it a go, also it has good characteristics for crossing)

So as of right now I have to order some swabs. Do q tips work just as well? Do I need to wrap in tin foil and pressure cook first?

I'll get a thread going with pics off the mystery strain (possible oakridge) and South American showing traits and then start there with trying a cross but having a mystery strain in the mix isn't really any good lol so I'll be waiting for these other strains to germ ofcourse.

Sorry not trying to hack your thread lol just kinda letting you know where I'm at since I haven't been posting on my projects or taking pics recently.

Will do that tho, start documenting. :)


Even if it ain't all good. It's all good enough.


*edit*

Not trying to jack your thread, I meant. Got a new phone and I gotta get used to it's stupid spell checks all over again.

Edited by DarkNchildlike, 03 April 2021 - 03:15 PM.

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#284 fahtster

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Posted 03 April 2021 - 03:14 PM

Cool. Yeah you definitely need to sterilize the qtips.. your plan sounds good.. a lot of ppl wrap a set in foil and then put them in a qt jar and sterilize a bunch at once and then leave it sealed u til you need one and grab from there and reseal to keep them as clean as possible

Faht
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#285 DarkNchildlike

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Posted 03 April 2021 - 03:17 PM

I also want to get some cholicine, or however its spelt, in the future.

Even if it ain't all good. It's all good enough.
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#286 DarkNchildlike

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Posted 03 April 2021 - 03:18 PM

Nice good idea, thanks faht!

Even if it ain't all good. It's all good enough.
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#287 fahtster

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Posted 03 April 2021 - 03:34 PM

I’d definitely use a well filtered lid (as in lots of FAE so that the foil packs are allowed to dry and stay clean). Maybe even a ring with just a couple pieces of tyvek or a decent sized filter disk.

Faht

#288 DarkNchildlike

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Posted 03 April 2021 - 04:00 PM

Word good looking out.

Even if it ain't all good. It's all good enough.

#289 fahtster

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Posted 04 April 2021 - 11:43 AM

PE/pfrs clone bag looking great
257415E1-C888-448B-8532-E68FB06628E4.jpeg

PE/RW cv/brf cakes look good and smell good.. getting excited for these
9649E97A-94B5-44CF-9B01-AF6F2054757D.jpeg

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#290 YoshiTrainer

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Posted 04 April 2021 - 12:15 PM

Looking great Faht!
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#291 pastyoureyes

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Posted 05 April 2021 - 09:54 AM


I also want to get some cholicine, or however its spelt, in the future.
Even if it ain't all good. It's all good enough.


For anyone interested in alternative methods of obtaining crosses, look into "yeast transformation mix". I am thinking this could possibly be adapted to crossing cubes but I don't have the time right now to test it out.
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#292 fahtster

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Posted 05 April 2021 - 09:10 PM

PE/pfrs suspect cross clone bag looking good..
F9C5BB67-305E-4063-B54D-B51114FC474A.jpeg B26DCAEE-1C76-415E-9449-101F6CEFA794.jpeg

I’m starting to think they’re the “flower” pheno I had of this bin.. https://mycotopia.ne...1612169193.jpeg (middle to the left)

They’re cool af looking but I’m pretty sure they were sporeless.. there’s a chance, albeit small, that they’re making clear spores (I doubt it) but its something I’m going to investigate this time if I don’t see pigmented gills since I was unaware that was even possible before but others have suggested it.

I was hoping for purple spores so I could share some here but I cloned the fruit when it was still a large pin so I didn’t get a chance to see it mature and which culture from that bin I had actually cloned. The way those caps are starting to get wavy, it’s not looking good even though it’ll be pretty neat and make for a nice pic

Faht

Edited by fahtster, 05 April 2021 - 09:17 PM.

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#293 fahtster

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Posted 07 April 2021 - 10:09 PM

Bit of a photo dump since these are so photogenic. Although it woulda been nice to get prints, I like the non spore drop :). But it’s also unfortunate I won’t be able to take these guys further.

6FEFDAE5-150E-45AB-A418-3AE0F6E6E390.jpeg 1A5C2D1C-E3D2-4045-B51B-7C3D387F674A.jpeg E4B975BA-EF66-40A3-AACA-18654613C986.jpeg
86997C9C-3B27-45F8-BD64-4BE4AE0D1D18.jpeg 166D5639-32F3-4521-B15E-FE56FA949615.jpeg

Faht

Edited by fahtster, 07 April 2021 - 10:09 PM.

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#294 fahtster

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Posted 08 April 2021 - 08:29 PM

Got some action happening on the PE/RW cakes. :).
74A21521-59DC-440A-B11E-37E6A08A9616.jpeg

Faht
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#295 fahtster

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Posted 08 April 2021 - 08:30 PM

Oops double post

Edited by fahtster, 08 April 2021 - 08:30 PM.


#296 rockyfungus

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Posted 08 April 2021 - 09:59 PM

Not seeing any whyteness, did you notice any? 


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#297 fahtster

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Posted 08 April 2021 - 10:48 PM

Not yet.. there’s some really light pins but I would’ve expected RW to pin before PE, so who knows.. super early though

Faht

#298 DarkNchildlike

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Posted 10 April 2021 - 12:53 AM

Faht, I just got germination on pfredspore lizardking maybe AA+ and alakabenzi,. Plus the mystery variety/ suspected oakridge clones are done.

I think I'm gonna get maybe four more varieties going and just try my hand at crossing everything like crazy.

I'll document ofcourse but its gonna be a month before everything is going..

I'm gonna try getting fresh wet sporulation of teo varieties like same day right next to each other and also try one variety wait a week then next variety.

Pastyoureyes I'll check that out as well!

Even if it ain't all good. It's all good enough.
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#299 rockyfungus

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Posted 10 April 2021 - 06:40 AM

If you look back a few pages I was hear hesitant about making a spore syringe, felt out of practice. Not only was it quite easy but the results were on par with my trusted agar.

Just spawned a tiny tester of AA+/PE, few more finishing up. Waiting on my 1st transfers to get large enough for grain.

Faht, how do you fruit tiny testers?

I was going to see how much room I had for casing(s) after it consolidates. Then fruit in the tupperware or put in a freezer bag with perlite/holes (as needed)


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#300 fahtster

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Posted 10 April 2021 - 08:41 AM

Faht, I just got germination on pfredspore lizardking maybe AA+ and alakabenzi,. Plus the mystery variety/ suspected oakridge clones are done.
I think I'm gonna get maybe four more varieties going and just try my hand at crossing everything like crazy.
I'll document ofcourse but its gonna be a month before everything is going..
I'm gonna try getting fresh wet sporulation of teo varieties like same day right next to each other and also try one variety wait a week then next variety.
Pastyoureyes I'll check that out as well!
Even if it ain't all good. It's all good enough.

Nice! Yeah it definitely takes a good amount of time but a lot of excitement builds up during the waiting and when you get the results you were looking for, the payoff is great. I know I’m excited to see what you do.

If you look back a few pages I was hear hesitant about making a spore syringe, felt out of practice. Not only was it quite easy but the results were on par with my trusted agar.
Just spawned a tiny tester of AA+/PE, few more finishing up. Waiting on my 1st transfers to get large enough for grain.
Faht, how do you fruit tiny testers?
I was going to see how much room I had for casing(s) after it consolidates. Then fruit in the tupperware or put in a freezer bag with perlite/holes (as needed)

Sounds like you’re finding what I did... swabs aren’t generally as dirty as we think. Letting the swabs dry for a week+ really helps, I think.

What size tester do you have.. is it a jar or small tray? I use ziplocks with perlite like you said, for small jars. I put the jars (2 or 3) in a bigger glad container... the 8 cup ones (they fit nicely inside the 2 gallon bags). then I spoon in wet perlite around the jars and then poke needle holes in the bag starting at the container top edge level, every couple of inches all the way to the top of the bag. I double up the amount of holes at the fruiting surface level like this , ‘ , ‘ , ‘ , ‘ in a band around the whole bag.

https://mycotopia.ne...1615235089.jpeg
https://mycotopia.ne...1613945880.jpeg

For larger testers that I use glad containers for, I just use whatever size ziplock they fit into snugly and do the same hole configuration. I think once you make a couple, you’ll figure out that you have a lot of options with ziplocks because of all the sizes they come in... you’ll get ideas. Just gotta keep the rh up and FAE balanced and they’ll fruit no problem. For a little tray like this, https://mycotopia.ne...1615196711.jpeg, I literally throw some wet perlite into the bottom of a ziplock (I think I used 1 gallon) and sit the tray on top of the perlite and poke holes the same way in the bag.

Here’s an inner stem poke clone that I did of a fruit off another tester jar of PE/crs. I’m pretty sure the cloned fruit was just PE that didn’t have the classic pheno but it’s hard to tell. A couple are petty blobby but then there’s the bigger fruits. There was another medium sized fruit that was dropping spores against the bag that I cut off because I thought the spores looked rusty.. turned out they weren’t and it was just bad lighting in the room I was in lol. These are a head scratcher and I don’t think I’m going to go further with them... I have other PE/crs that are displaying both characteristics more but these testers are so easy to throw together and fruit that it’s worth it just to check out and see what fruits.
832271DB-F415-49A0-A0D7-91361B856040.jpeg CB5D750C-F766-4F39-BBC5-21D880E990FC.jpeg

For even larger tester (1 quart spawn grows) I love the 2 gallon ziplocks sitting in 8 cup containers that I’m sure you’ve seen.. I’m going to be doing a write up of them as soon as the last few I have fruiting finish up. I recently figured out that if I get a lot of side pins, because of how much bigger the 2 gallon bag is than the sub block, I can simply take the bag out of the 8 cup container (it’s on the outside of the bag and used to just form the sub into a block shape while it colonizes.. there’s a pic a couple posts up actually so you probably do know what I’m talking about lol) and pull the bag away from the sub sides and the fruits will grow normally and it just becomes a block sitting inside a 2 gallon bag and there’s lots of room on all sides. Here’s a pea+ f1 clone I have fruiting rn..

8BF0396B-26FA-4F83-AFFF-15BC4B7FF257.jpeg

You can see the 2 fruits on the front short side... those are side pins. That’s actually one of the quarts of the revived original pea+ clone that I dried out since last November.. more info on that here https://mycotopia.ne...dpost&p=1479937

Faht

Edited by fahtster, 10 April 2021 - 08:54 AM.

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