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Re: AGAR ENVY! (Anything and All things agar!) [Re: anders] * 1
    #29320050 -



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"For the powerful freedom-takers who rob justice from us all, that's why I keep my pistol loaded, powder, cap, and ball..."
Read more, post less!
...turns out, I'm an amateur at everything

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Re: AGAR ENVY! (Anything and All things agar!) [Re: KlarnEvil9] * 1
    #29320055 -



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I've got skills, they don't know where to use me
I'm like the best dressed guy at the nude beach
Nice to meet you!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: ThurmanMurman] * 2
    #29320057 -

ThurmanMurman said:
Im starting to think every user here is a carefully crafted Catnip puppet and all of this is happening just to troll me.


I'm the only one not out to get you, man.


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

∴Genetics Info∴Ps. ovoideocystidiata and Native Americans∴

∴The Official Woodlovers Thread∴Outdoor Mushroom Cultivation∴

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Re: AGAR ENVY! (Anything and All things agar!) [Re: catnip40] * 2
    #29320060 -

Catnip's hand has been up my ass. That makes me a puppet right?


--------------------
I've got skills, they don't know where to use me
I'm like the best dressed guy at the nude beach
Nice to meet you!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PuffM4gicDr4gon]
    #29320062 -

:whatev:


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The Hitchhiker’s Guide to the Shroomery - BeefSupremeJr
A Love Letter to New Growers - Smellyhobbit

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #29320075 -

How many spores?


Edited by Puffin (08/20/25 05:04 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Puffin]
    #29320124 -

Puffin, be sure to spread it out.

I would like to understand what went wrong with my Subtrop cultures.

T - 7 August, on 11 August:


12 August:


And then before I know it, it's all mold on 20 August:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: anders] * 1
    #29320130 -

anders said:
Puffin, be sure to spread it out.


I refuse to do it!

I actually did just do a bunch of spread plates using some of those spreader stick things but those were serial dilutions to isolate monokaryons, ~25 CFUs per plate.

This super concentrated drop in the middle is for fun :laugh:


--------------------
Protoplast Fusion


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Puffin] * 3
    #29320204 -

Puffin said:
How many spores?




are we guessing? are you going to count them? do i win something if i guess right?

im guessing... 450,000 spores


--------------------
The Hitchhiker's Guide to the Shroomery
Visit our cultivation museum
The Hitchhiker's guide Pt. 3

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Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefExtreme]
    #29320210 -

IOW, you're not really going to learn much from this Puffin.  You'll get several puffballs wadded together, and intermingling myc.

It is adviseable to spread out genetics so you can choose the most promising and isolate the nasties.

But, we all learn our own ways.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: anders] * 3
    #29320246 -

theres no way to tell whats promising on a plate. 
its like looking over tokyo and deciding whos promising.

take a few neighborhoods and send em.  then see what buidings rise.  there is your promise.

ive sent many germ plates to grain, whole.  all of tokyo is a city full of promise.


--------------------
The Hitchhiker's Guide to the Shroomery
Visit our cultivation museum
The Hitchhiker's guide Pt. 3

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Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefExtreme] * 1
    #29320255 -

BeefSupremeJr said:
are we guessing? are you going to count them? do i win something if i guess right?

im guessing... 450,000 spores


The clumps settled at the bottom of a test tube. I pipetted them and then held the pipette downwards so the clumps gathered at the tip, and inoculated the agar with a single ~0.05ml black drop. I reckon that the black drop was 10% spores by volume, which would be approximate 7 million spores.

So not quite all of Tokyo, but getting there!



--------------------
Protoplast Fusion


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Puffin]
    #29320263 -

Puffin said:
BeefSupremeJr said:
are we guessing? are you going to count them? do i win something if i guess right?

im guessing... 450,000 spores


The clumps settled at the bottom of a test tube. I pipetted them and then held the pipette downwards so the clumps gathered at the tip, and inoculated the agar with a single ~0.05ml black drop. I reckon that the black drop was 10% spores by volume, which would be approximate 7 million spores.

So not quite all of Tokyo, but getting there!




fascinating.  do you have your own thread? id like to follow along.  im interested to see what youre up to.  also i cant have you going around unrated pardner


--------------------
The Hitchhiker's Guide to the Shroomery
Visit our cultivation museum
The Hitchhiker's guide Pt. 3

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PuffM4gicDr4gon]
    #29320281 -

PuffM4gicDr4gon said:
Catnip's hand has been up my ass. That makes me a puppet right?


wait until you all find out catnip is my puppet.
:yeahthatsright:


--------------------
:hypnotoad:
Making Liquid Culture :  Go insane with LC
Mushboy likes these teks : Making shroom tea
Dank Education

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Re: AGAR ENVY! (Anything and All things agar!) [Re: mushboy]
    #29320285 -

Puppetception


--------------------
I've got skills, they don't know where to use me
I'm like the best dressed guy at the nude beach
Nice to meet you!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PuffM4gicDr4gon] * 1
    #29320290 -

:mindblown:


--------------------
:hypnotoad:
Making Liquid Culture :  Go insane with LC
Mushboy likes these teks : Making shroom tea
Dank Education

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Re: AGAR ENVY! (Anything and All things agar!) [Re: anders] * 3
    #29320316 -

anders said:
Puffin, be sure to spread it out.

I would like to understand what went wrong with my Subtrop cultures.

T - 7 August, on 11 August:


12 August:


And then before I know it, it's all mold on 20 August:



I mean, I'm just another noob, but when you were already cutting chunks out of your plate on the 12th, it doesn't seem surprising that 7 days later it was mostly mold?

A lot of experienced folks earlier in the thread seemed insistent that trying to cut contam out of a plate was a losing battle. This seems like pretty strong evidence of that being correct...


--------------------
:blackspy:00010010
"When a long train of abuses and usurpations, pursuing invariably the same Object evinces a design to reduce them under absolute Despotism, it is their right, it is their duty, to throw off such Government, and to provide new Guards for their future security."

The Hitchhiker's Guide to the Shroomery <-- This is the way.



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Re: AGAR ENVY! (Anything and All things agar!) [Re: Synack] * 2
    #29320323 -

:rofl:


--------------------
It doesn't matter what i think of you...all that matters is clean spawn
I'm tired do me a favor

I'M A MEMBER FIRST, TC WHEN NEEDED AND MOD WHEN I HAVE TO BE !!!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: cronicr]
    #29320325 -



--------------------
I've got skills, they don't know where to use me
I'm like the best dressed guy at the nude beach
Nice to meet you!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefExtreme] * 3
    #29320358 -

Quote:
BeefSupremeJr said:fascinating.  do you have your own thread? id like to follow along.  im interested to see what youre up to.  also i cant have you going around unrated pardner


My ambition is to create novel hybrids using protoplast fusion. For the challenge and adventure more than anything.

The first stage is to isolate monokaryotic cultures for as many Psilocybe species as possible. I started with subaeruginosa and natalensis today; creating spore solutions, counting spores under the microscope, doing serial dilutions to make a solution with ~25 CFUs per 0.1ml (single spores or clumps is how I'm defining that), and spreading it onto plates. I'll be watching these for single spore germinations and doing early transfers, then proving monokaryotic status with a microscope. I've got spores for hispánica, ovoideocystidiata, cyanescens, azurescens and cubensis up next.

The stage after this is to generate auxotrohpic strains for each monokaryotic culture using UV mutagenesis. Auxotrophs are mutants that can't create a certain nutrient so rely on it being in their environment to survive. The idea is that each parent has a different auxotrophism, then after protoplast fusion (forced mating essentially), when the resulting cultures are cultured on minimal media, the only ones that will survive and grow are hybrids, with the working gene from the other parent for to fix their auxotrophism. Self mated parents do not survive do not survive, so it's a way to find 'the needle in the haystack' when a hybridisation does occur.

I've started writing it up in my shromery journal but it's all very early days and most things are super draft. This will be the main thread at the moment:
Isolating Monokaryotic Cultures, but it is part of a bigger series.

I'll write up today's process in there at some point tomorrow and start using that as a log.

I can DM you any time anything notable happens, if you're interested?


--------------------
Protoplast Fusion


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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