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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Wanna Suggest Some Sectors?
    #25533452 -

First Agar work, hence the stupid labeling, condensation, so apologies in advance for the errs.

I've been doing my transfers based off the 16 guidelines for sectoring from TMC so far,  but I'd love to hear from those more experience on which they'd go from next! Also, the dishes with debris are MAE dishes that have not been PC'ing well.
The individual pictures are the most recent transfers photographed on 10/12. All of these are 3 transfers away from MS , with the last dish 4 from some grain spawn. Included the large group picture of assorted earlier transfers if anyone can think of use for them at this point.
  • Agar 1:



  • Agar 2:



  • Agar 3:



  • Agar 4:



  • Agar 5:



  • Agar 6:



  • Agar 7:



  • Previous Transfers:



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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25533612 -

Shit dude when you got that much going on I'd say just pick one section of each plate with your preferred growth pattern, you're not gonna know how it performs until you grow each one out anyway. What exactly are you after?


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Re: Wanna Suggest Some Sectors? [Re: Noisecontrol]
    #25534190 -

I have 6 varieties I'm working with, and while working on my isolation technique I'm interested in seeing if any at all observable/distinctive strain Phenotypic variation can be observed persistently from each given controlled environments.
If a cube is a cube, and overarching characteristics (rhizomorphism, linear hyphal growth, density etc.) are universal, I'd like to see a convergence between them all. If there are some variations, albeit just within the isolations I have since ya know millions of strains out of one spore print, that show reason to keep them (faster colonizing, pin set, etc) I'd think that'd be all the more interesting.
It's all probably futile, I'm sure someone here could tell me why, but at the end of the day its fun and I can get could practice/improved technique.


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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25534293 -

Here's Today

  • Agar 1:



  • Agar 2:



  • Agar 3:



  • Agar 4:



  • Agar 5:



  • Agar 6:



  • Agar 7:



    *Not Previously Posted*
  • Agar 8:



  • Agar 9:



  • Previous Transfers:



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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25535119 -

You'll find that it takes a lot of effort to get true isolates because through the process of elimination, the sectors can remingle and you must be diligent to keep them separate. Also, most cultures that perform well are compiled of different strain combinations, and cutting down on the combinations may sacrifice the qualities you're after. Just something to be aware of. If you do end up seeing this all the way through to true isolates please share them as they are fascinating. Great looking plates tho. Any way I can help with your condensation and debris?


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Re: Wanna Suggest Some Sectors? [Re: Noisecontrol]
    #25539538 -

Any suggestions for LME? I bought local and it just  seems to be the same through the bag. The moisture in the plates is just gonna take more precision on my end.


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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25540444 -

Keeping the LME airtight is necessary as over time it sucks up moisture and becomes clumpy, and when that happens it doesn't dissolve as well. Add the agar and LME when your water is cold and bring to a boil. I personally boil for about 5 minutes, sometimes more depending on the state of my LME when I use it. And tho these things help, I've found that LME is generally problematic because of this. You can boil significantly and decant the sediment but it's a tedious extra step that people typically don't want to add. Dextrose is a good alternative, same with potato water or even more simply is water from soaking/cooking your grain. You can freeze it and thaw as needed. Hope this helps.

P.s. if you pour your agar around 120F you won't have condensation. That or you can stack plates however I find it easier to not stack when working in a SAB. The stacking channels the heat upward to the top plate (which will have condensation unless you put another container of hot liquid like a mug of hot water on top). You can put your agar container in a water bath and temp the water to get it to a workable temp. If it solidifies, 185F will melt it, and solidification happens at 105F. You wanna be as close to that 105 as possible while still having enough time to finish all your pours. I like to keep my thermometer in the bath water and watch the temp after I remove my agar and start pouring, this way I have an idea as to what temp I'm pouring at and how much time I have.


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Re: Wanna Suggest Some Sectors? [Re: Noisecontrol] * 1
    #25540467 -

i get crystal clear(almost) LME agar by mixing cold and pcing right away. why boil agar twice?


also got a batch of lme thats over a year old and it hasnt clumped yet.. grow more shrooms so it doesnt hang around would be good advice:tongue:


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Re: Wanna Suggest Some Sectors? [Re: mushboy]
    #25540790 -

First, Noisecontrol thank you for the feedback, it's much appreciated and I'll keep posting updates on the successive transfers. So to circumvent the LME I actually used beet root powder since I had around the house. I figured it would be a two birds one stone by also adding color to the media (it wasn't the color disappeared after being PC'd). Second, I used an IR thermometer at the time of pouring the agar, however still had a few with condensation. I have to try water baths again, but they didn't make too much different previously.

Also Mush, whats your time PC'd? No sediment? And yes I need to grow more, getting some wbs prep started tomorrow  :paperairplane:


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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25541070 -

It usually takes me at least 10 transfers before I start to see really defined sectoring.
I take transfers from every individual sector (I might leave out obviously weak looking sectors). I aim to take transfers from at least ten sectors from one culture,sometimes as many as twenty.

Often times the next plates will still show some sectoring and they will need another transfer or too further before they are monocultures.

I then take those 10-20 monocultures, label them + fruit from each plate. If I got lucky there will be a keeper to slant.


How many transfers along from spore germination are those cultures? It looks like they have a way to go before defined sectoring will show. There are so many strains present and crowded together at this stage that you won't see any lines of seperation


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Amateur Mycologists United
AMU Q&A

Edited by mushpunx (10/16/18 03:46 AM)

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Re: Wanna Suggest Some Sectors? [Re: mushboy]
    #25541140 -

mushboy said:
i get crystal clear(almost) LME agar by mixing cold and pcing right away. why boil agar twice?


also got a batch of lme thats over a year old and it hasnt clumped yet.. grow more shrooms so it doesnt hang around would be good advice:tongue:



I didn't think it would blend that well skipping the preboil but those look great. I assume you still pour, mush?


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Edited by Noisecontrol (10/16/18 05:14 AM)

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Re: Wanna Suggest Some Sectors? [Re: Noisecontrol]
    #25541149 -

Sometimes I have to heat up my agar before I sterilize it because my LME clumps as it gets older.
But I just stick my media bottles in the microwave for a couple minutes.
With fresh LME I don't usually have to do that


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Amateur Mycologists United
AMU Q&A

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Re: Wanna Suggest Some Sectors? [Re: mushpunx]
    #25542071 -

Here are the same Transfers as of today, looking a little despondent/translucent... my speculation as to why is such:

1. New media, which has the mycelium "inspecting" it
2. The media is too hard, nutritious, both, or lack thereof?
3. Weak mycelium from poorly selected sectors
4. Contaminated or simply contaminants

The potential sectors I see are selected, if anyone sees a reason for others, one specifically, none at all and should just start over please let me know.


  • Agar 1:



  • Agar 2:



  • Agar 3:



  • Agar 4:



  • Agar 5:



  • Agar 6:



  • Agar 7:




  • Agar 8:



  • Agar 9:



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Re: Wanna Suggest Some Sectors? [Re: mushpunx]
    #25542089 -

Mushpunx I do feel I'll need to transfer further as well. I am, as another option, considering to wait and let them grow further, if 1. is the suspected issue and I simply need to see stronger growth. The concern with this option for me though is letting the myc interweave and botch isolation.


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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25542295 -

Rowboat said:
Mushpunx I do feel I'll need to transfer further as well. I am, as another option, considering to wait and let them grow further, if 1. is the suspected issue and I simply need to see stronger growth. The concern with this option for me though is letting the myc interweave and botch isolation.


Right now there are so many strains present you can't find any sectors. Sometimes cultures actually look like monocultures early on because of this even!

But the more they are transfered (take tiny slivers BTW, rice sized), the more strains are getting cut out.
Eventually you will start seeing lines of separation and individual sectors.

I like to let my cultures grow half dollar size or so to look for them.

Im not sure what you mean about myc interweaving, the sectors will fan out separate from each other, you'll be able to see the lines of separation.
You'll often get more than one sector by accident , you just separate them out on the next plate.


--------------------

Amateur Mycologists United
AMU Q&A

Edited by mushpunx (10/17/18 01:16 AM)

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Re: Wanna Suggest Some Sectors? [Re: mushpunx]
    #25543118 -

1, 3, and 6 look promising. Myc will act different each time its food changes. Dont worry everything looks great


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Re: Wanna Suggest Some Sectors? [Re: mushpunx]
    #25543309 -

I was referring to This advice RR gave a while back.

RogerRabbit said:
Deej3987 said:
I want more space for the mycelium to colonize maybe sectoring would be easier?? I don't know, just something I want to try. Where did you say you got yours at?


If the mycelium colonizes more, the various strains will tend to merge into a coherent whole, defeating the purpose of isolating.  I use three section dishes, and never allow the mycelium to grow larger than a half-dollar.  This way, individual strains can be isolated and tested.
RR


It's from a while back, but nonetheless what I've gone with for transfers so far. If your saying to let them intersect further, I'll do transfers once further grown out as well, I did however bring the size of the transfer down to rice sized for the first few I did this evening :thumbup:


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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25543568 -

He is just saying not to let your cultures fill out the plate too much yea

That's what I do too, I'll let my cultures grow to about half dollar size, smaller than that and the sectors haven't usually fanned out much yet. If it isn't showing sectors yet I just make a transfer from a healthy spot.


--------------------

Amateur Mycologists United
AMU Q&A

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Re: Wanna Suggest Some Sectors? [Re: mushpunx]
    #25579050 -

Just updating, switched some of the media to: Maltodextrose, Yeast, activated Charcoal. There were some I did keep the same, but seeing some interesting variation. Completed the majorities' 6th transfers and SAT's 7th this afternoon. Also, some test jars with some very appealing growth.


SAT Agar



Mazatepec Agar




KSSS Agar




PEU Agar



APE Agar



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Re: Wanna Suggest Some Sectors? [Re: Rowboat]
    #25579152 -

The SAT tho. Currently working on 4 different lines with it, such aggressive myc n so beautiful. Been testing their fruiting capabilities and I even found some genes that don't fruit. Have you run into this yet?


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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