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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: Isolate Shortcut Experiment - *updated with pics [Re: Breakfast Crew]
    #12597956 -

has anyone here tried using just two spores on agar before because that would be all you needed right.

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Re: Isolate Shortcut Experiment - *updated with pics [Re: drewb149]
    #12598271 -

drewb149 said:
has anyone here tried using just two spores on agar before because that would be all you needed right.


That is why I recommended the microscope. I think directly putting 2 spores on agar would be very hard so I think starting two spores under a microscope slide and waiting for them to connect would be the first process, then transfer that to a dish.
I do a lot of my syringes to agar and have good results if they are somewhat fresh, though it is a good way to try and get old ones going also.


--------------------
By the livin' Gawd that made you,
    You're a better man than I am, Gunga Din

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Re: Isolate Shortcut Experiment - *updated with pics [Re: Breakfast Crew]
    #12598325 -

I don't mean to discourage you, but it might be simpler and more effective to use clone tissue or a young pin. Might take you maybe 1 extra step to isolate it?

What you are doing is just a tissue culture. If you're going to do tissue culture to agar, why not do it simply and easily.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: Isolate Shortcut Experiment - *updated with pics [Re: anonjon]
    #12598905 -

anonjon said:
I don't mean to discourage you, but it might be simpler and more effective to use clone tissue or a young pin. Might take you maybe 1 extra step to isolate it?

What you are doing is just a tissue culture. If you're going to do tissue culture to agar, why not do it simply and easily.



I prefer complicated and failure-prone!  :tongue2:

No, actually I just wanted to see if I could get to an isolate from one transfer, but you are absolutely right.


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Re: Isolate Shortcut Experiment - *updated with pics [Re: Breakfast Crew]
    #12599232 -

Breakfast Crew said:
IMO, spores should never be directly transferred to agar. Using live tissue or colonized grain is a good starting point.


That is absolutely incorrect.  You inoculate agar with spores so you can isolate strains as they emerge.

To the OP, taking colonized grains isn't an isolation technique.  The strains have most likely already combined into a single organism by the time the jar is fully colonized.  Non-compatible strains might exist in a grain jar, but unlikely on the same grain.

You got rhizomorphic mycelium on the coir due to the structure and moisture content.  Mycelium often goes from rhizo to cottony, etc., depending on substrate, regardless of whether it's an isolate or not.

To isolate strains, you separate sectors, not necessarily based on the mycelium type.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Isolate Shortcut Experiment - *updated with pics [Re: RogerRabbit]
    #12600208 -

RogerRabbit said:
Breakfast Crew said:
IMO, spores should never be directly transferred to agar. Using live tissue or colonized grain is a good starting point.


That is absolutely incorrect.  You inoculate agar with spores so you can isolate strains as they emerge.

To the OP, taking colonized grains isn't an isolation technique.  The strains have most likely already combined into a single organism by the time the jar is fully colonized.  Non-compatible strains might exist in a grain jar, but unlikely on the same grain.

You got rhizomorphic mycelium on the coir due to the structure and moisture content.  Mycelium often goes from rhizo to cottony, etc., depending on substrate, regardless of whether it's an isolate or not.

To isolate strains, you separate sectors, not necessarily based on the mycelium type.
RR


Thanks for the clarification!  Guess I can't be lazy after all.  What is your take on the agar picture - do you see sectors or is that likely to be single strain?


--------------------
:mushroom2:Bulk Grow Substrate Calculator Spreadsheet - Download :heartpump: Thread:mushroom2:
*Downloaded over 3,500 times!* Also try the DMT Changa Mix Calculator

Stonesun's Amazing Sclerotia/Stones Guide:mushroom2:Vasodilator List:mushroom2:Niacinamide - reduce trip anxiety


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Re: Isolate Shortcut Experiment - *updated with pics [Re: gornyhuy]
    #12600237 -

You could probably answer that yourself if you counted how many times you transferred. I think his dvd said like 5 times at least? It's been a while since I watched it though.


--------------------
By the livin' Gawd that made you,
    You're a better man than I am, Gunga Din

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Re: Isolate Shortcut Experiment - *updated with pics [Re: Breakfast Crew]
    #12601478 -

Breakfast Crew said:
IMO, spores should never be directly transferred to agar. Using live tissue or colonized grain is a good starting point.


I put spores on agar just to make sure the print is clean.


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


My spawn lab
Using hand blender as eberbach
Mycomaster 2000
Few prints for trade

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Re: Isolate Shortcut Experiment - *updated with pics [Re: madi]
    #12618787 -

Here is a nice set of the agar growth from start to finish.
Is this single strain?
Obviously I dropped a little tiny myc over on the side during the transfer, otherwise it would be a perfect bulls eye pattern.



What are the concentric rings indicating? Is that day/night growth?


--------------------
:mushroom2:Bulk Grow Substrate Calculator Spreadsheet - Download :heartpump: Thread:mushroom2:
*Downloaded over 3,500 times!* Also try the DMT Changa Mix Calculator

Stonesun's Amazing Sclerotia/Stones Guide:mushroom2:Vasodilator List:mushroom2:Niacinamide - reduce trip anxiety


Edited by gornyhuy (05/24/10 08:37 AM)

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Re: Isolate Shortcut Experiment - *updated with pics [Re: gornyhuy]
    #12619013 -

gornyhuy said:
What are the concentric rings indicating? Is that day/night growth?


I want to know too.


--------------------
You make it all possible. Doesn't it feel good?

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Re: Isolate Shortcut Experiment - *updated with pics [Re: Doc_T]
    #12619041 -

Quote:
What are the concentric rings indicating? Is that day/night growth?


Day/night as well as temperature variations will cause that.  If you're attempting to isolate, you shouldn't let the dish grow out that much.  All the strains will consolidate into a single organism via anastomosis.  Strain isolation needs to be conducted early.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Isolate Shortcut Experiment - *updated with pics [Re: RogerRabbit]
    #12619067 -

Question re:

RogerRabbit said:
  If you're attempting to isolate, you shouldn't let the dish grow out that much.  All the strains will consolidate into a single organism via anastomosis.  Strain isolation needs to be conducted early.
RR


If it consolidates into one organism, it's actually a true isolate then, right?
Not necessarily the best of the dish, but usable in controlled experiments, for example?


--------------------
You make it all possible. Doesn't it feel good?

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Re: Isolate Shortcut Experiment - *updated with pics [Re: Doc_T]
    #12619608 -

Doc_T said:
Question re:

RogerRabbit said:
  If you're attempting to isolate, you shouldn't let the dish grow out that much.  All the strains will consolidate into a single organism via anastomosis.  Strain isolation needs to be conducted early.
RR


If it consolidates into one organism, it's actually a true isolate then, right?
Not necessarily the best of the dish, but usable in controlled experiments, for example?


Good question.  I'm apparently skilled in sterile/agar stuff enough to do the work, but I have some fundamental knowledge/technique gaps.  What should I read to remedy that? Stamets?  RR's DVDs?  I've never seen a step by step pictorial on strain isolation or cross breeding with venom or anything else 'advanced'...


--------------------
:mushroom2:Bulk Grow Substrate Calculator Spreadsheet - Download :heartpump: Thread:mushroom2:
*Downloaded over 3,500 times!* Also try the DMT Changa Mix Calculator

Stonesun's Amazing Sclerotia/Stones Guide:mushroom2:Vasodilator List:mushroom2:Niacinamide - reduce trip anxiety


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Re: Isolate Shortcut Experiment - *updated with pics [Re: Doc_T]
    #12621655 -

Doc_T said:
Question re:

RogerRabbit said:
  If you're attempting to isolate, you shouldn't let the dish grow out that much.  All the strains will consolidate into a single organism via anastomosis.  Strain isolation needs to be conducted early.
RR


If it consolidates into one organism, it's actually a true isolate then, right?
Not necessarily the best of the dish, but usable in controlled experiments, for example?


It's one organism in the sense that it can transmit signals and nutrients and water. But maybe it won't be genetically homogenous? i.e. some genetic variety in the nuclei getting passed around?

just speculating. It does make sense that over time the various similar substrains might consolidate.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: Isolate Shortcut Experiment - *updated with pics [Re: RogerRabbit]
    #12621845 -

Sorry about mistake. I should have said clarified my reason for that statement and how starting from live tissue is faster than starting from spores.

Quote:
If you're attempting to isolate, you shouldn't let the dish grow out that much.  All the strains will consolidate into a single organism via anastomosis.  Strain isolation needs to be conducted early.

  That I did not know. Learn something new everyday! :lolsy:


--------------------
$30 Stir plates
Agar's Grain Liquid Culture Tek
How to case

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Re: Isolate Shortcut Experiment - *updated with pics [Re: Breakfast Crew]
    #12621926 -

Quote:
IMO, spores should never be directly transferred to agar. Using live tissue or colonized grain is a good starting point.


Are you serious? Are you suggesting that inoculating agar with a spore syringe or print to agar is a bad idea?

Quote:
o the OP, taking colonized grains isn't an isolation technique.  The strains have most likely already combined into a single organism by the time the jar is fully colonized.  Non-compatible strains might exist in a grain jar, but unlikely on the same grain.

You got rhizomorphic mycelium on the coir due to the structure and moisture content.  Mycelium often goes from rhizo to cottony, etc., depending on substrate, regardless of whether it's an isolate or not.



I knew he was going to say that.....I did.

I think its agar that is a very nutritious medium for the mycelium so therefore if the strain isn't the most robust it'll be like "hey this is more than enough, I'll be cottony and chow down on whats here" but if its a real winner it will say "I know this is more than enough BUT I'm a thug, enough is never enough must go find moooorrreeeee, I'm so rhiiiizzooooooo"

Thats how it goes.

Grains and bulk are less nutritious and are prone to cause the mycelium to be rhyzo regardless, never minding the exceptions to the rules ie, extremely cottony strains, environmental conditions, etc.


--------------------
:three:

Edited by camplo (05/24/10 07:12 PM)

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Re: Isolate Shortcut Experiment - *updated with pics [Re: camplo]
    #12623171 -

dude grains and bulk, ie. compost/manure, are two of the most if not the most nutritious substrates to grow mushrooms on.

thats why we grow with these materials.

Edited by drewb149 (05/24/10 10:24 PM)

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Re: Isolate Shortcut Experiment - *updated with pics [Re: drewb149]
    #12623928 -

Bulk is not that nutritious..it doesnt need to be sterilized so you can do large amounts at once. Thats why we use bulk. Also because less nutritious things colonize faster. Thats why as stated strains tend to always look rhizo on bulk. Not that I havnt seen cottony stuff on manure..


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay

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Re: Isolate Shortcut Experiment - *updated with pics [Re: scatmanrav]
    #12623953 -

scatmanrav said:
Bulk is not that nutritious..it doesnt need to be sterilized so you can do large amounts at once. Thats why we use bulk. Also because less nutritious things colonize faster. Thats why as stated strains tend to always look rhizo on bulk. Not that I havnt seen cottony stuff on manure..


:themoreyouknow:


--------------------
By the livin' Gawd that made you,
    You're a better man than I am, Gunga Din

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Re: Isolate Shortcut Experiment - *updated with pics [Re: scatmanrav]
    #12624727 -

scatmanrav said:
Bulk is not that nutritious..it doesnt need to be sterilized so you can do large amounts at once. Thats why we use bulk. Also because less nutritious things colonize faster. Thats why as stated strains tend to always look rhizo on bulk. Not that I havnt seen cottony stuff on manure..



if im wrong im wrong but this is where i gathered my info maybe i was presumptive.

why compost is better than other substrates

sorry scroll to the top to read the main post not sure if it directs you to the top or bottom of the thread

Edited by drewb149 (05/25/10 07:30 AM)

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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