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InvisibleLeviticus969
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Re: Lab access [Re: SloppyJoseph]
    #23759288 - 10/21/16 09:29 PM (7 years, 3 months ago)

I'd rather not have my questions answered by smart alecs that think I'm stupid

Quote:

You don't need a geneticist to tell you this.




Quote:

Alright fucknut, ask the geneticist.  Sorry I bothered correctly answering your stupid ass question.
:banyouverymuch:




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InvisibleLeviticus969
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Re: Lab access [Re: irequirechocolate]
    #23759294 - 10/21/16 09:30 PM (7 years, 3 months ago)

Quote:

irequirechocolate said:
Hopefully this will satisfy both of you: asexual spores are genetically identical to the parent. But, because of epigenetics and environmental cues, you're still not going to have an identical mushroom.
EDIT: I have not reached the level of taking prints yet; I'm still waiting on my first set o'cakes. But maybe eventually I can extract DNA from a mother than daughter and sequence the two. This should not be hard to do.




Awesome response. Thanks man and yes, it would be interesting to see the mother and daughter relation.


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InvisibleInocuole
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Re: Lab access [Re: Leviticus969] * 2
    #23759328 - 10/21/16 09:43 PM (7 years, 3 months ago)

If the literal fact that you do not need a geneticist to tell you how mushroom spores work upset you, how do you propose you'll get anything done in life?  And it was true, wasn't it?  Because geneticist here is talking about asexual spores.  Mushroom spores can have many sexes, and not all are inter-compatible.  A single spore does not produce the mycelium we're looking for, therefore not identical to parent.  They were right about epigenetics being a factor though.  :thumbup:

Frankly your insulting my effort to inform you, and calling me a know it all and being an ungrateful little prick should be considered a lot more offensive than my simply telling you that certain information is not necessarily geneticist level knowledge.  Anyone who's successfully grown and participated in the community here would know these things.  The point of being here is to talk about those things.  If you just now waited for the opportunity to ask a geneticist, then that means you weren't using this site to its potential, which is the real shame here.  If you'll ignore the correct answer in favor of the answer you want to hear a la appeal to authority, then I do indeed regret even bothering to share that information.

But you know, I actually do have a life, and I'm not interested in wasting it explaining myself to someone so thin skinned, so if you want to play the victim and act like you didn't walk right into everything I threw at you, then by all means.



--------------------
                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

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Edited by Inocuole (10/21/16 09:49 PM)


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OfflineSloppyJoseph
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Re: Lab access [Re: Inocuole]
    #23759348 - 10/21/16 09:50 PM (7 years, 3 months ago)

Quote:

Inocuole said:
If the literal fact that you do not need a geneticist to tell you how mushroom spores work upset you, how do you propose you'll get anything done in life?  And it was true, wasn't it?  Because geneticist here is talking about asexual spores.  Mushroom spores have many sexes, and not all are inter-compatible.

Frankly your insulting my effort to inform you, and calling me a know it all and being an ungrateful little prick should be considered a lot more offensive than my simply telling you that certain information is not necessarily geneticist level knowledge.  Anyone who's successfully grown and participated in the community here would know these things.  The point of being here is to talk about those things.  If you just now waited for the opportunity to ask a geneticist, then that means you weren't using this site to its potential, which is the real shame here.  If you'll ignore the correct answer in favor of the answer you want to hear a la appeal to authority, then I do indeed regret even bothering to share that information.

But you know, I actually do have a life, and I'm not interested in wasting it explaining myself to someone so thin skinned, so if you want to play the victim and act like you didn't walk right into everything I threw at you, then by all means.




And if that's tldr for you Leviticus...

:shutthefuckup:

And continue to start shit with respected cultivators, and take the advice of someone who's never taken a spore print on what spores are like. See how much longer you last around here.


--------------------
AMU Q&A


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Invisibleirequirechocolate
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Re: Lab access [Re: Inocuole]
    #23759390 - 10/21/16 10:05 PM (7 years, 3 months ago)

Quote:

Inocuole said:
Quote:

irequirechocolate said:
Hopefully this will satisfy both of you: asexual spores are genetically identical to the parent. But, because of epigenetics and environmental cues, you're still not going to have an identical mushroom.





The spores aren't asexual though.




Despite my career, it's been a super long time since I took a bio class  or anything that focused on the life cycle of fruiting fungi, so there are some major gaps in my knowledge. Still, the definition of a fungal spore is pretty much that: the asexual reproductive body of a fungus.
This is actually kind of a heavy topic that I was going to start to get into, but instead just look at this. It's hella basic: http://bioweb.uwlax.edu/bio203/2011/bielmeie_luke/Reproduction.htm

Anyway, it's not really an issue. I'm just trying to work efficiently using what I have to get some nice and bulky results w/out having to buy spores over and over. (This is why I grow my own yeast for brewing as well).

After jumping around though, it looks like what I need to do is let my current cakes fruit, clone a fatty and then let it grow on some rich media agar plate. Unless you've got a protocol for this and/or how to move this over to a FC I will go back to mining old threads :wink:


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InvisiblePastywhyteMDiscord
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Re: Lab access [Re: irequirechocolate] * 2
    #23759418 - 10/21/16 10:13 PM (7 years, 3 months ago)

Imperfect fungi are asexual. However we don't want to be growing those. Nasty. Cubes of course are higher fungi and not only do they require two spores to exchange genetic information they also need to be compatible with each other.

As far as potency and genetics go spores are essentially a reset. While some well stabilized lineages have tendencies, the true potential they have is (assuming no serious genetic degredation of the lineage) ultimately only limited by the genome of the species. As has been said many times, isolate and use agar to get consistent results. Anyone denying that is simply too lazy to put in the work and like all half ass cultivators will see much disappointment.


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Invisibleirequirechocolate
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Re: Lab access [Re: Pastywhyte]
    #23759510 - 10/21/16 10:42 PM (7 years, 3 months ago)

Found this on a super old thread, https://www.shroomery.org/forums/showflat.php/Number/15621268:

Quote:

mellowparty said:
I inoculated some cube spores in YPD liquid media and left the flask on a shaker at 30degrees constant temperature. They germinated in a few days producing spherical colonies reminiscent of some LC contaminants (they produced mushrooms so no they were not contams)

I also inoculated some cube spores in YPD liquid and left them in a test tube at room temperature (around 20degrees). It took them longer to germinate and the mycelium they formed was not spherical colonies but irregularly shaped mass of mycelium. Then I used a vortex machine to break it apart and used that for mushrooms. Worked fine.




This would basically be routine practice for me in the lab. I'm going to try in out but I'm curious as to what might be the following steps once germination in liquid culture happens. Unfortunately the original poster was banned years ago...


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OfflineNDStepp84
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Re: Lab access [Re: irequirechocolate]
    #23759529 - 10/21/16 10:52 PM (7 years, 3 months ago)

Lol @ putting spores in LC
:goodluckwiththat:


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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Invisibleirequirechocolate
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Re: Lab access [Re: NDStepp84]
    #23759558 - 10/21/16 11:04 PM (7 years, 3 months ago)

I may go in to lab over the weekend and set this up. We should know w/in a couple days.
If I get growth I will use it to inoculate some jars at the same time as I use plain spores on others. Could be an interesting race.


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OfflineNDStepp84
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Re: Lab access [Re: irequirechocolate]
    #23759560 - 10/21/16 11:06 PM (7 years, 3 months ago)

You don't need a lab to put dirty spores into sugar water, you can do that shit at home.


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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OfflineSloppyJoseph
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Re: Lab access [Re: irequirechocolate]
    #23759561 - 10/21/16 11:07 PM (7 years, 3 months ago)

Spore in LC is the worst way to germinate them. Use agar if you have a lab... or if you don't.


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AMU Q&A


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Invisibleirequirechocolate
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Re: Lab access [Re: SloppyJoseph]
    #23759571 - 10/21/16 11:12 PM (7 years, 3 months ago)

Quote:

NDStepp84 said:
You don't need a lab to put dirty spores into sugar water, you can do that shit at home.



Sure I could, but the lab has the added benefit of pre-made media, sterile flasks & tubes, incubators, shakers, roller drums, an autoclave for the jars, etc.
We'll see wha happens.
Quote:

SloppyJoseph said:

Spore in LC is the worst way to germinate them. Use agar if you have a lab... or if you don't.



I'll try it on some plates while I'm there.


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OfflineSloppyJoseph
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Re: Lab access [Re: irequirechocolate]
    #23759578 - 10/21/16 11:15 PM (7 years, 3 months ago)

ND is trying to say it doesn't matter if your lab is sterile, the spores are dirty. At home you can make all the sterile media yourself too.


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OfflineNDStepp84
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Re: Lab access [Re: irequirechocolate]
    #23759588 - 10/21/16 11:18 PM (7 years, 3 months ago)

What you don't get is no syringe or print is clean, spores are dirty it dosen't matter what you have if you inject contaminations into it, you can't look at an LC and tell if it's clean or not. That's what agar is for, you grow the culture on a 2d surface and transfer away from any contaminations. When done properly an LC is inoculated with a clean agar wedge, colonized then tested on agar. 
Edit:whathesaid:

Check the date of the info on this site, use the advanced search feature to limit your search results to the last few years


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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Invisibleirequirechocolate
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Re: Lab access [Re: NDStepp84]
    #23759603 - 10/21/16 11:26 PM (7 years, 3 months ago)

Quote:

SloppyJoseph said:
At home you can make all the sterile media yourself.



I don't doubt it, but in lab the media has already been made, saving me time & monies. There's cabinets full of liters and liters and stacks and stacks of plates.
Quote:

NDStepp84 said:
What you don't get is no syringe or print is clean, spores are dirty it dosen't matter what you have if you inject contaminations into it, you can't look at an LC and tell if it's clean or not. That's what agar is for, you grow the culture on a 2d surface and transfer away from any contaminations. When done properly an LC is inoculated with a clean agar wedge, colonized then tested on agar. 
Edit:whathesaid:

Check the date of the info on this site, use the advanced search feature to limit your search results to the last few years




I gotchya. I can isolate from agar and move it to LC if needed.
If the syringe comes w/ contaminated spores, why do people have such success just inoculating right into a cake jar w/out this being a concern?


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OfflineNDStepp84
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Re: Lab access [Re: irequirechocolate]
    #23759617 - 10/21/16 11:32 PM (7 years, 3 months ago)

BRF cakes are more forgiving compared to grain, and especially a low oxygen nutrient rich liquid solution. PF tek is a beginners method to get a few mushrooms, a cake contams no big deal throw it out, you have a bunch more. You have 5 quarts of grain spawned to bulk and one is bad you can loose everything.


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


Edited by NDStepp84 (10/21/16 11:42 PM)


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OfflineSloppyJoseph
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Re: Lab access [Re: NDStepp84]
    #23759625 - 10/21/16 11:37 PM (7 years, 3 months ago)

Use the shit out of those plates then. It would be awesome to have access to unlimited agar like that. Just don't waste your time making an LC with spores when you have all that.


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Invisibleirequirechocolate
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Re: Lab access [Re: NDStepp84]
    #23759632 - 10/21/16 11:41 PM (7 years, 3 months ago)

Quote:

NDStepp84 said:
BRF cakes are more forgiving compared to grain. PF tek is a beginners method to get a few mushrooms, a cake contams no big deal throw it out, you have 5 quarts of grain spawned to bulk and one is bad you can loose everything.




Ah, I see. I haven't had any contam problems yet so I hadn't considered that the source would be from the syringe's contents. I assumed if someone had an issue it was due to improper sterilization of the jar and substrate.

Hey, while I've got your attention, what kind of grain do you use? I frequently home brew and have always felt like there must be a good way to re-use the malt grain after steeping it. It's still got a ton of nutritional content. I've had a buddy make bread w/ it before, but perhaps I could use it for this...


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OfflineNDStepp84
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Re: Lab access [Re: irequirechocolate]
    #23759647 - 10/21/16 11:46 PM (7 years, 3 months ago)

Most of us use whatever is cheapest/easily sourced with the hull still intact, whole oats, rye berries, wheat, wild bird seed preferably with no/low cracked corn.


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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OfflineSloppyJoseph
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Re: Lab access [Re: NDStepp84]
    #23759653 - 10/21/16 11:48 PM (7 years, 3 months ago)

Malted barley would probably get too starchy to be useful for this. Nutritional content doesn't really matter, actually the richer it is the slower the mycelium will colonize.


--------------------
AMU Q&A


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