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My agar trials - Quest for rizomorphic


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#1 rhapsodyclaymore

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Posted 16 May 2021 - 04:34 AM

Hi everyone,
 
Another member thought it best to move to here since my original thread has grown out of the new members area.
 
1 week ago, I nocc'ed up 8 plates with a GT spore syringe (fresh purchased) and yesterday, after looking through the plates, only 1 seemingly had the best section to make a transfer from compared to the others. I made 3 transfers into new cups.
 
The source sample. 3 small cuts taken near the top with the best rizo strands there.
before-transfer.JPG
 
After the transfer yesterday. The cups have embossed lids and the light source incorrectly casts dark sections on the agar that do not exist at all. All the cups are the same color on the surface.
after-transfer.JPG
 
At 4am (after waking up in the middle of the night), the cuts weren't doing very much, but now some 7 hours later, they have fuzzed up and are spreading out on the new agar.
 
I've ordered some more 53ml jars coming this week (28x) and I will clean up some older agar jars (i poured too much in) and make a new fresh batch that's the same in the blue cups you see there.
 
I have 5 more of the original 2oz batch cups I've created initially which I'll use for further transfers from the 3 pictured in this thread. I think I'll eventually have a good batch before long.
 
A question to everyone: Once I have transferred some good rizo stuff, the best way to keep a constant supply of this strain would be to hold a cup back (make extra transfers) from the ones going into spawn jars to keep cloning this into new agar, pretty much creating an infinite supply of this good rizo? Does this make sense? Or would I need to go back to my spore syringe and work up that way?

 



#2 sandman

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Posted 16 May 2021 - 07:15 AM

Hi there it's your friendly neighborhood sandman here. I'm going to say some negative things, but I'm just trying to help you.

 

Ease up on the blue dye. Too much dye is probably making your cultures grow wonky. I recommend to lose the dye and use activated charcoal powder. 

 

I am not a fan of these tiny cups. They are so tiny that there isn't much room for your transfers to grow. Plus your fingers are so close to the edges when you take the lid off. I don't like that. I recommend these style with a no-pour tek.

 

You can keep the culture going by holding some mother plates back in the fridge and taking a small cut out to grow a new plate or LC out for each grow.


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#3 rhapsodyclaymore

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Posted 16 May 2021 - 08:31 AM

Hi there it's your friendly neighborhood sandman here. I'm going to say some negative things, but I'm just trying to help you.

 

Ease up on the blue dye. Too much dye is probably making your cultures grow wonky. I recommend to lose the dye and use activated charcoal powder. 

 

I am not a fan of these tiny cups. They are so tiny that there isn't much room for your transfers to grow. Plus your fingers are so close to the edges when you take the lid off. I don't like that. I recommend these style with a no-pour tek.

 

You can keep the culture going by holding some mother plates back in the fridge and taking a small cut out to grow a new plate or LC out for each grow.

Hey! Actually, the criticism is absolutely fine. And yes, the blue coloring was from my very first agar batch, and I will admit, I used too much. I'm not into baking, so I didn't realize how concentrated that stuff was. I'll work on getting some bigger cups. I plan on doing another batch of no-pours this week.

 

Do you have an EU friendly recommendation? The cups (4 meds) are around 30 euros here. Astronomical in cost.


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#4 sandman

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Posted 16 May 2021 - 08:43 AM

I am sure you have a local equivalent at your large grocery store. The twisting lids are preferred. Snapping motion can cause flaking in your SAB.



#5 rockyfungus

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Posted 16 May 2021 - 08:44 AM

Another note, rhizo is overrated. Just make sure it's obviously myc. No need to isolate homogenous looking growth, you're just limiting the chances of having fruitable genetics IMHO.


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#6 rhapsodyclaymore

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Posted 16 May 2021 - 08:49 AM

I am sure you have a local equivalent at your large grocery store. The twisting lids are preferred. Snapping motion can cause flaking in your SAB.

5179zdV65FL._AC_SL1333_.jpg

Height approx. 5.2 cm diameter approx. 4.5 cm

I've actually got a bunch of these 53ml ones. They are smaller than what you've suggested, but this is what I have right now. Twist lock lids. These may be a bit small tho. You suggested bigger ones for letting it grow out more. I assume that these wouldn't be that useful.

 

As an alternative, I can get these 230ml twist off preserve jars pretty cheap. They have quite the wide mouth which would make it great to put into spawn jars. Height: 7.1 cm. More wider base than tall which is good.

81KXpgjbe4L._AC_SL1500_.jpg

 

Thoughts?


Edited by rhapsodyclaymore, 16 May 2021 - 09:02 AM.


#7 rhapsodyclaymore

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Posted 16 May 2021 - 08:50 AM

Another note, rhizo is overrated. Just make sure it's obviously myc. No need to isolate homogenous looking growth, you're just limiting the chances of having fruitable genetics IMHO.

Gotcha, I'll keep that in mind when I get some stuff up and running better.



#8 UnHeisenbug

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Posted 16 May 2021 - 11:36 AM

Another note, rhizo is overrated. Just make sure it's obviously myc. No need to isolate homogenous looking growth, you're just limiting the chances of having fruitable genetics IMHO.


That’s really interesting. Did you learn this experimentally or from some source?

#9 rhapsodyclaymore

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Posted 16 May 2021 - 11:43 AM

 

Another note, rhizo is overrated. Just make sure it's obviously myc. No need to isolate homogenous looking growth, you're just limiting the chances of having fruitable genetics IMHO.


That’s really interesting. Did you learn this experimentally or from some source?

 

I mean, the one thing I can say about cleaning up or letting it go wild is, from research, you get more consistently grown caps rather than a jungle of stuff. It's still good, I'm sure, but you'll have a lot of different growths and stuff.



#10 sandman

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Posted 16 May 2021 - 12:36 PM

Those red 230ml ones should do great for no-pour agar. I'd put a polyfil wad in a hole in the lid for a filter. 


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#11 bezevo

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Posted 16 May 2021 - 01:07 PM

( Good idea sandman ) OP  could get fancy and ad an SHIP (self healing injection port ) or mae one with regular clear silicone no need to by RTF ....        FANCY



#12 rockyfungus

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Posted 16 May 2021 - 01:07 PM

 

Another note, rhizo is overrated. Just make sure it's obviously myc. No need to isolate homogenous looking growth, you're just limiting the chances of having fruitable genetics IMHO.


That’s really interesting. Did you learn this experimentally or from some source?

 

I used to isolate and try and get the monoculture or whatever people are shooting for with that agar porn. Tested out multiple "isolates" some overlayed like crazy and never fruited. Others just side-pinned or had sparse pinning. Don't think it's my conditions. After 20 different strains and maybe a few 1,000s of plates I'm confident as others have seen with MSS syringes, the closer to a germ plate the better chance of a "canopy" or better fruits to clone from. 

Also, if you've poured enough plates at different thickness with different media you'd also know rhizo almost switches back to tomentose or whatever depending on variables I can't control for the most part. There's also a tomentose gang out there...

 

Look at Fahts work with rubbing different strains together, he's going direct from sporeload. 


Edited by rockyfungus, 16 May 2021 - 01:12 PM.


#13 rhapsodyclaymore

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Posted 16 May 2021 - 02:15 PM

Those red 230ml ones should do great for no-pour agar. I'd put a polyfil wad in a hole in the lid for a filter. 

Great! Order placed (thank you amazon). I need to see how many I can fit into my PC. We don't have the super tall ones like you have in the states. Mine wasn't cheap either (300 euros). I need to see when they arrive how many I can stack vertically.


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#14 sandman

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Posted 16 May 2021 - 02:30 PM

Yea no pours are best when you are only working with few plates. They take up a lot more space than regular petris. A little pc would make it a bitch to do a lot.


Edited by sandman, 16 May 2021 - 02:32 PM.


#15 rhapsodyclaymore

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Posted 16 May 2021 - 03:44 PM

Yea no pours are best when you are only working with few plates. They take up a lot more space than regular petris. A little pc would make it a bitch to do a lot.

Mine is large by EU standards and is 10L (dimensions are 40.5 x 31.5 x 25.5 cm) @ max of ~13psi (a bitch to find PC-ers here that can do that much. 15psi is the north american standard). It's not too bad, tbh. The benefit of no-pour is i can do 2 batches of PC'ed jars without much problem. It's just another PC session.



#16 UnHeisenbug

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Posted 17 May 2021 - 04:34 AM

Yea no pours are best when you are only working with few plates. They take up a lot more space than regular petris. A little pc would make it a bitch to do a lot.

Mine is large by EU standards and is 10L (dimensions are 40.5 x 31.5 x 25.5 cm) @ max of ~13psi (a bitch to find PC-ers here that can do that much. 15psi is the north american standard). It's not too bad, tbh. The benefit of no-pour is i can do 2 batches of PC'ed jars without much problem. It's just another PC session.

There is an Indian made pressure cooker called Hawkins Bigboy. I picked mine up for about a hundred euros and is 10L. That’s the cheapest one I’ve found that is 15psi in the EU.
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#17 rhapsodyclaymore

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Posted 17 May 2021 - 07:36 AM

Yea no pours are best when you are only working with few plates. They take up a lot more space than regular petris. A little pc would make it a bitch to do a lot.

Mine is large by EU standards and is 10L (dimensions are 40.5 x 31.5 x 25.5 cm) @ max of ~13psi (a bitch to find PC-ers here that can do that much. 15psi is the north american standard). It's not too bad, tbh. The benefit of no-pour is i can do 2 batches of PC'ed jars without much problem. It's just another PC session.
There is an Indian made pressure cooker called Hawkins Bigboy. I picked mine up for about a hundred euros and is 10L. That’s the cheapest one I’ve found that is 15psi in the EU.
I opted in the end for a high end Swiss made one (can't remember the brand) just because I might want to use it also for food too. Figured it was an investment. It's really high quality and I can fit 12x 250ml jars inside for making grain spawn. I need to see when my agar jars come tomorrow how many I can fit inside.

Short tiny update on the growth in the 3 agar cups. Growth is very good. Good rizomorphic growth that looks pretty healthy. I'll make transfers from these into the new jars once there is sufficient growth. It is tons easier to see growth with 1 point of growth rather than ~10 different points which got really confusing. I'm still learning, but so far things are not too bad.

Edited by rhapsodyclaymore, 17 May 2021 - 08:19 AM.


#18 rhapsodyclaymore

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Posted 18 May 2021 - 03:11 AM

Ok, so after 4 days, they are looking quite too.

 

1.JPG

 

This one looks slower than the rest. The reason why is because I couldn't get the transfer chunk with the myc side anywhere close to the plate and it was the complete opposite. The myc had to take over the chunk first before going to the rest of the plate which it is doing right now.

2.JPG

 

3.JPG

 

So far so good. Looks pretty healthy.

 

New jars arrive today and I'll make a batch of new plates in prep for the transfers from the already growing plates.


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#19 rhapsodyclaymore

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Posted 18 May 2021 - 12:13 PM

Ok, I've got one batch of jars on the stove in the PC going. I've got 14 jars inside and 11 packed up and ready to go once that batch is done. I'm using unmodified lids and wrapping them with foil, cracking the lid slightly open to avoid pressure buildup. Worked fine for the 28x 53ml jars I did, so I'm going to run with that again.

The growth has gone maybe 0.5mm bigger in the 3 cups in the pics above. Probably in another 2 days I'll need to do some transfers to the new agar plates to let them really grow.

I did use less food coloring this time. I'm learning. And I used blue since the white myc on blue agar plates is super pretty.

Oh, when I post another update on those 3 agar cups, after those pictures were taken today, some additional white myc spots started opening up on one of the cups. The reason is when I was checking them out late Saturday night, I accidentally dropped one of them and the chunk of agar flipped and stuck to the wall of the agar cup. So, after some bumping around, I managed to get it mostly back in the center again, however, it touched a few areas of the agar plate and the growth from those touches is now starting to appear. Just an FYI.

Edited by rhapsodyclaymore, 18 May 2021 - 12:22 PM.


#20 sandman

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Posted 18 May 2021 - 02:01 PM

Can you get activated charcoal powder? It's good stuff to add to agar. Better than food color.






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