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Check this out, B+ on brf/water(agar substitute) fruiting from jar.


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#1 Billcoz

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Posted 03 January 2019 - 12:08 AM

 Yeah, I made these a couple months ago and transferred some sector wedges to real agar and wanted to see if they would fruit from the jar. I was thinking it would have to be very aggressive genetics to want to fruit from a 1/4" deep brf/water "puck".

 

 I am about to grab the giant pin from the jar(see pics) and clone it, it should be very clean as it in a jar with micropore and that is in a sealed baggie, and it still fruited.

 

 So my thought is that if I clone this thing to agar, then make an LC, it should give aggressively fruiting genetics as it actually fruited in the sealed bagged jar, is this a good plan?

 

 Check it out, you can kind of see the big pin in one jar, it has a tiny head(cap) but the pic don't show it-IMG_20190102_185735.jpg IMG_20190102_185336.jpg IMG_20190102_185402.jpg IMG_20190102_185508.jpg IMG_20190102_185528.jpg IMG_20190102_185652.jpg IMG_20190102_185535.jpg IMG_20190102_185236.jpg -You can kinda see the arching pin.

 

 Thumbnails are the second jar with no pins, So what do you guys think, would the pin be good to put straight into an LC, or would you put it on agar, then make LC? Remember it's been sealed in the clean jar(beige color in jars is brf) for two months.

 

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#2 coorsmikey

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Posted 03 January 2019 - 12:34 AM

Three weeks to fruit on agar is aggressive. That's a good way to select aggressive strains. Two months isn't aggressive at all. Clone that bad boy and put it back to BRF and in two months, maybe less with oxygen you will have fruits. What do you think that the immune system of something that has been in a bubble? Try the same experiment with future nutrients and some of the competitors that it will face later? I used to subscribe to the same thinking but have changed my mind along the way.



#3 Billcoz

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Posted 03 January 2019 - 01:02 AM

Three weeks to fruit on agar is aggressive. That's a good way to select aggressive strains. Two months isn't aggressive at all. Clone that bad boy and put it back to BRF and in two months, maybe less with oxygen you will have fruits. What do you think that the immune system of something that has been in a bubble? Try the same experiment with future nutrients and some of the competitors that it will face later? I used to subscribe to the same thinking but have changed my mind along the way.

It's been sealed up and the pin has been in there for A MONTH!. It's weird because it has not opened at all, the cap is tiny, I thought it was just myc until I saw the lilcap today, but the fruit grew in less than a month after I made em, why would it not mature as fast?

 

 Are you saying that it's immune system might be too weak, and it will die when exposed? I was gonna open it in the SAB and put a tiny piece on agar, and a piece into LC, but would it be better to hgrow it out on agar before LC?

 

 How would I introduce competitors, just exposure? 



#4 Billcoz

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Posted 03 January 2019 - 02:52 AM

 Okay I re-read your reply coorsMikey and I understand what you were saying now, I was confused by the "future nutes" thing, I have some agar ready and I'm gonna work on it in the next couple days and I'll post pics of what IO get.

 

 I got this idea, after being frustrated with jars i used for agar dishes, it's hard to see any sectors that aren't really obvious because the lid is in the way, even with a light shining through. 

 

 I got these fancy-assed 1/4 pt jars that are shorter than the normal wide-mouths with agar in em, and when I go to inoculate the agar in the SAB, I'll take the jar lids off, put the tissue or spores in the center, then slide the whole jar into a new ziplock baggie(they are sterile on the inside), and then screw the lid ring on over the baggie. That way I can see the shit.

 

 Another cool thing I tried is, instead of food coloring, I added the ink from a neon highlighter marker to help "highlight" contams. It is water soluble and non-toxic, so I figured it wouldn't hurt, but it was an old, dried up marker and I had to squeeze the shit out of the ink-soaked foam thing inside just to get a couple drops to come out.

 

I haven't inoculated any yet, check it oot-IMG_20190103_024123.jpg IMG_20190103_023155.jpg IMG_20190103_023020.jpg IMG_20181230_171633.jpg

 Since the ink is water-soluble, when it wa still liquid the water, being on top of the agar, had most of the ink, so it looks separated, with the color of the agar coming through, I let the jagar cool and harden with the lids off before pcing, so there is almost no condensation.


Edited by Billcoz, 03 January 2019 - 02:56 AM.

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#5 Billcoz

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Posted 08 January 2019 - 03:22 AM

 I transferred some pieces from each of my old agar cultures(cubes; B+, Burma), to these "highlighter" agar jars. They were showing slight growth after 24 hrs. What does everyone think of my baggie tek, to make it easy to see the myc/contams, and the highlighter idea?-

 

IMG_20190105_190437.jpg IMG_20190107_110004.jpg IMG_20190107_110055.jpg

 

 I also have some pink(supposed to be red) agar made up that I am using some other experiments, and starting some Blue Meanies(cube) and P. Tampanensis, from prints.

 

 I still have yet to take the tissue/clone from the pin in the brf/h2o jar, it's still looking alive in the there. I will be doing that this morning.  It has been sealed under 3 layers of micropore and also in a ziplock baggie.

 

 Would it be good enough to just reach my xacto knife into the jar and cut a piece from the base of the pin, or would it still be better to pull the whole pin, then peel it apart and shave a sample?


Edited by Billcoz, 08 January 2019 - 03:23 AM.


#6 Deleena24

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Posted 08 January 2019 - 12:02 PM

Love the baggie idea! Amazing nobody else has thought of something so simple.

Edited by Deleena24, 08 January 2019 - 12:03 PM.


#7 Billcoz

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Posted 09 January 2019 - 04:10 AM

2 days after transferring, 3rd sector tranfer from the original brf jar, hope it's a good fruiting isolate, I haven't fruited this genetic sector out yet. 

IMG_20190109_034927.jpg IMG_20190109_034753.jpg IMG_20190109_034653.jpg

I am pretty sure it's not a single-genetic sector yet, maybe 1 or 2 more transfers, what do y'all think? 

 

How likely is someone to get genetic isolates that do not fruit?

 

Remember, I am doing this agar stuff to get the technique down, I will be selecting genetics based on fruiting, this is just playing around to see how it works and how i can manipulate things.

 

I realize I need to get better at taking smaller tissue samples for tranfers.


Edited by Billcoz, 09 January 2019 - 04:12 AM.


#8 Deleena24

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Posted 09 January 2019 - 11:18 PM

Bill, I thought you dont have a flowhood...how the hell are you doing such good agar work?
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#9 Billcoz

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Posted 12 January 2019 - 05:58 AM

Well, on one of the "highlighter" agar jars, there is some tiny satellite colonies showing up. I am pretty sure it is just myc(it looks like myc, it's white) because it is right where I had dropped the tiny tissue sample on the edge of the agar plate and tried to pick it back up a few times, bouncing it around a bit, to get it in the middle of the dish.

 

Is this a problem? Is this likely? I will watch for any color changes until I can do more sab work and get some more sectors transferred to new agar. They will surely be isolated to single genetic sectors by the next transfer. The rhizo-growth pattern is beautiful, just like a spore print. I consider it "myco-art".



#10 Billcoz

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Posted 12 January 2019 - 10:04 AM

Bill, I thought you dont have a flowhood...how the hell are you doing such good agar work?

Geez, ha, now I'm embarrassed, but this is how-

IMG_20190105_145738.jpg -And-IMG_20190112_092715.jpg -And check my cool torch-IMG_20190112_095242.jpg

Agar has not been difficult for me, but I am new still. It is intimidating the first couple times, but it gets easy. 


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#11 Deleena24

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Posted 12 January 2019 - 12:47 PM

Yeah I used a couple plates in my SAB/GB, but it is cumbersome. I was able to clone cleanly, but how do you pour it? Is there a tek which allows you to do it all in the PC?

My LFH got lost in shipping somehow, so I am still waiting, but I'd still like to get some good work done while I'm waiting for it.
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#12 Billcoz

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Posted 13 January 2019 - 09:33 AM

Yeah I used a couple plates in my SAB/GB, but it is cumbersome. I was able to clone cleanly, but how do you pour it? Is there a tek which allows you to do it all in the PC?

My LFH got lost in shipping somehow, so I am still waiting, but I'd still like to get some good work done while I'm waiting for it.

I use the "no pour" agar tek from this tek @ shrmry, but jars are better than the lil tupperware dishes he shows, they are too light and ca be tippy. 

 

You just mix any kind of agar and put it in the jars while hot and still liquid, then let it harden & cool a bit before putting the lid on to keep condensation down, & I put folded paper towel on the lid to catch any water before it can get in, then tin foil, and pc for 40-45 min.


Edited by Billcoz, 13 January 2019 - 09:33 AM.

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#13 JACKOLANTERN

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Posted 13 January 2019 - 09:47 AM

 Okay I re-read your reply coorsMikey and I understand what you were saying now, I was confused by the "future nutes" thing, I have some agar ready and I'm gonna work on it in the next couple days and I'll post pics of what IO get.

 

 I got this idea, after being frustrated with jars i used for agar dishes, it's hard to see any sectors that aren't really obvious because the lid is in the way, even with a light shining through. 

 

 I got these fancy-assed 1/4 pt jars that are shorter than the normal wide-mouths with agar in em, and when I go to inoculate the agar in the SAB, I'll take the jar lids off, put the tissue or spores in the center, then slide the whole jar into a new ziplock baggie(they are sterile on the inside), and then screw the lid ring on over the baggie. That way I can see the shit.

 

 Another cool thing I tried is, instead of food coloring, I added the ink from a neon highlighter marker to help "highlight" contams. It is water soluble and non-toxic, so I figured it wouldn't hurt, but it was an old, dried up marker and I had to squeeze the shit out of the ink-soaked foam thing inside just to get a couple drops to come out.

 

I haven't inoculated any yet, check it oot-attachicon.gifIMG_20190103_024123.jpgattachicon.gifIMG_20190103_023155.jpgattachicon.gifIMG_20190103_023020.jpgattachicon.gifIMG_20181230_171633.jpg

 Since the ink is water-soluble, when it wa still liquid the water, being on top of the agar, had most of the ink, so it looks separated, with the color of the agar coming through, I let the jagar cool and harden with the lids off before pcing, so there is almost no condensation.

Hey Bill I wonder what you will see under a black light with the highlighter agar if it will be picked up into the myc. It is Fluorescein and is used as a tracer dye.


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#14 Billcoz

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Posted 13 January 2019 - 09:56 AM

 

Hey Bill I wonder what you will see under a black light with the highlighter agar if it will be picked up into the myc. It is Fluorescein and is used as a tracer dye.

 

I didn't think of that, I'll have to find a black light. I don't think I own one.


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#15 Billcoz

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Posted 15 January 2019 - 08:17 PM

Deleena24, here's proof that I'm not so great at agar yet, I bounced the tissue sample around tryin to get it to the center of the dish, so there's all sorts of satellite colonies around the big one. I think there are still sectors here, this is the most isolated I;ve seen any, but I think I could get a more uniform plate, IDK-IMG_20190115_180823.jpg IMG_20190115_180848.jpg IMG_20190115_180740.jpg IMG_20190115_181007.jpg Anyone know if Im close to a single sector yet?



#16 Deleena24

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Posted 16 January 2019 - 01:59 PM

Deleena24, here's proof that I'm not so great at agar yet, I bounced the tissue sample around tryin to get it to the center of the dish, so there's all sorts of satellite colonies around the big one. I think there are still sectors here, this is the most isolated I;ve seen any, but I think I could get a more uniform plate, IDK- IMG_20190115_180823.jpg IMG_20190115_180848.jpg IMG_20190115_180740.jpg IMG_20190115_181007.jpgAnyone know if Im close to a single sector yet?


Hey, you've been able to pour and experiment with most of your dishes performing well and without contam. That's damn good for someone who just started and doesnt have a LFH, IMO.

It's nice to know you're the humble type, though. :)
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#17 Billcoz

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Posted 19 January 2019 - 12:46 AM

Thanks Deleena24, I need some confidence right now, I'm feeling like my grows are struggling

I'll probably just put all these agar dishes on popcorn, or make an LI syringe for brf, accept for one that is, I think, an isolated genetic strain, which I'll transfer a sample to new agar, then dump the rest of the plate on popcorn to fruit as a test. If it's anything special I might try to do genetics work, by taking prints and starting the selection process over, repeating, and gradually, selectively, reducing the genes, eventually getting something unique, that maybe I could call my own strain, & name it "BC+"(get it?).

 

Agar is fun, but you really gotta do TONS of transfers to keep a library. That's why I want to get a good genetic representation of each strain, down to where the fruiting results are predictable and unique, and have some master cultures of each, that way I don't have to make a hundred friggin agar plates a month. 

 

I'll post new pics in the next coupla days.


Edited by Billcoz, 19 January 2019 - 12:49 AM.


#18 jkdeth

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Posted 19 January 2019 - 08:27 AM

Lol. The strain game! I'd feel guilty renaming a named strain. I do however have a few prints labeled Unknown Cubensis. Growing it out now. I'm going to pick a phenotype, work it till its dominant and call it Unknown Henson. Cause that's what I think every time I see the label.
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#19 Billcoz

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Posted 21 January 2019 - 03:35 AM

Lol. The strain game! I'd feel guilty renaming a named strain. 

Lol, it's still like B+, but now it's BC+. I imagine it will take many cycles from spore to clone to spore etc to get to the point where it is a stable strain, but I feel like that's one of my goals in this hobby, to make my own unique cube. Maybe not actually rename something, but make my own version of a strain.



#20 jkdeth

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Posted 21 January 2019 - 09:20 AM

Just info gleaned from the "cloud" on myco knowledge, you should be able to stabilize a phenotype in about 6 cycles (12 grows) but take that with a grain of salt.
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