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OfflineErik77johansson77
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Agar isolation and fruiting (ALL)
    #22480480 - 11/05/15 10:21 AM (8 years, 2 months ago)

Hey guys,

I've learned from my mistakes. Did spore on agar and isolated to mono culture.

And did grow it out monotub (three times!). But it didn't give me any fruits. I thought I was doing something wrong. Which I did obviously. After a while I read that some cultures doesn't fruit. *suck*

That was disappointment for me, spent many months for nothing!.


My question is, this has to be a lot of agar plates right?

        A1
          |
A2  -          A2
|                  |
A3-A3    A3-A3
|        |      |        |

etc etc... end of the line after 6-8 transfers you get mono culture.

But that makes many monotubs to grow on to see if the isolates fruit.


My question is:
How do people do, grow in little boxes or what?


--------------------
/Erik


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InvisibleMad Season
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Re: Agar isolation and fruiting (ALL) [Re: Erik77johansson77]
    #22480539 - 11/05/15 10:35 AM (8 years, 2 months ago)



--------------------
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InvisibleSupalemonhaze
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Re: Agar isolation and fruiting (ALL) [Re: Mad Season]
    #22480747 - 11/05/15 11:31 AM (8 years, 2 months ago)

Its interesting to note that he started with a clone in that thread. I guess even when working with a clone fruiting is not a guarantee.

Kudos for the link. Was a nice read :takingnotes:


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OfflineDrCrumbs
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22480840 - 11/05/15 11:54 AM (8 years, 2 months ago)

Nice link, does he ever state what the yield was from the original MS?


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OfflineErik77johansson77
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Re: Agar isolation and fruiting (ALL) [Re: Mad Season]
    #22480906 - 11/05/15 12:12 PM (8 years, 2 months ago)

Quote:

Mad Season said:
http://www.shroomery.org/forums/showflat.php/Number/18219379





Thanks seen that. But still, who has time to make all of that?

Is this the thing people do when they say "You have to grow it, to see if it fruits" do ALL of them do monotub to check if their isolation has potential?

I was thinking it would be easier, sort of small batch or something.


--------------------
/Erik


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InvisibleSupalemonhaze
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Re: Agar isolation and fruiting (ALL) [Re: Erik77johansson77]
    #22481115 - 11/05/15 01:05 PM (8 years, 2 months ago)

Yes you can see if an isolate yeilds well even on PF cakes. I guess if it were me I would do mini monotubs first to see how it goes. PF cakes actually take more time to colonise than grains IME.


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InvisibleLocN9ne
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22481118 - 11/05/15 01:07 PM (8 years, 2 months ago)

Muda bottles work great for testing isolates.


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


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OfflineErik77johansson77
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22481128 - 11/05/15 01:09 PM (8 years, 2 months ago)

Quote:

Supalemonhaze said:
Yes you can see if an isolate yeilds well even on PF cakes. I guess if it were me I would do mini monotubs first to see how it goes. PF cakes actually take more time to colonise than grains IME.




It's agar, how do you transfer it to pf cake?

It's terrifying to think to make so many monos to get an isolation. And that's just to see if it fruits. 

It looked like PussyFartMs all mono did fruit (even if few was lost to trich)


--------------------
/Erik


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InvisibleKalistis
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Re: Agar isolation and fruiting (ALL) [Re: Erik77johansson77]
    #22481167 - 11/05/15 01:23 PM (8 years, 2 months ago)

You can turn agar in LC for injection to PF cakes.


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


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Offlineguyute22
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22481173 - 11/05/15 01:25 PM (8 years, 2 months ago)

Quote:

Supalemonhaze said:
Its interesting to note that he started with a clone in that thread. I guess even when working with a clone fruiting is not a guarantee.

Kudos for the link. Was a nice read :takingnotes:




i don't think that rate of non-fruiting is normal.  I think it was because it was PE.


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


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InvisibleSupalemonhaze
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Re: Agar isolation and fruiting (ALL) [Re: Erik77johansson77]
    #22481180 - 11/05/15 01:26 PM (8 years, 2 months ago)

You can make an LI or LC out of it if you got dry verm barrier. LI seems like you best choice though for fast results. Dont forget to transfer a piece to a new plate before you add the water though so if it fruits well you can keep it.


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InvisibleMad Season
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22481184 - 11/05/15 01:28 PM (8 years, 2 months ago)

Quote:

Supalemonhaze said:
Its interesting to note that he started with a clone in that thread. I guess even when working with a clone fruiting is not a guarantee.

Kudos for the link. Was a nice read :takingnotes:



That was the point to an extent. But if you don't isolate and just keep running with the clones multiple genetics, you should see much better shit. It seems genetic variation is actually a great thing. The fruits you desired were like that because of the multiple genetics. At least that seems to be the speculation.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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InvisibleSupalemonhaze
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Re: Agar isolation and fruiting (ALL) [Re: Mad Season]
    #22481231 - 11/05/15 01:46 PM (8 years, 2 months ago)

Genetic variety is the way a species survives after all. You know I just figured out how selfish that makes us. Isolating 1 strain for better fruiting. Just one strain all by its lonesome. Just so we can say "omg itsa canopy!" :p


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Offlinemushpunx
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Re: Agar isolation and fruiting (ALL) [Re: Supalemonhaze]
    #22482182 - 11/05/15 05:52 PM (8 years, 2 months ago)



This is why you don't want to just end up with one isolate from a culture.

You should keep transferring until you see sectoring (like in pic) and then take a transfer from *each* sector and then keep making transfers on each untill you have a whole bunch of isolates.

You should aim to end up with at Least 10 isolates from a culture to test out. If you just have one you have nothibv to compare it to, and unless your lucky it might be garbage. Usually 1 out of 10 is a keeper


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

Amateur Mycologists United
AMU Q&A


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OfflineYaMoonSun
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Re: Agar isolation and fruiting (ALL) [Re: mushpunx]
    #22482190 - 11/05/15 05:54 PM (8 years, 2 months ago)

Hundreds of petri dishes later :lol:


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Offlinemushpunx
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Re: Agar isolation and fruiting (ALL) [Re: YaMoonSun]
    #22482214 - 11/05/15 06:00 PM (8 years, 2 months ago)

Yea dude! Considering if you are working on 10 isolates you burn thru a sleeve  of plates every other transfer


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

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OfflinePsilocyBen17
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Re: Agar isolation and fruiting (ALL) [Re: YaMoonSun]
    #22482220 - 11/05/15 06:01 PM (8 years, 2 months ago)

put a gram or two of grain or sawdust ontop of an agar plate and it will fruit after colonizing.


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OfflinePsilocyBen17
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Re: Agar isolation and fruiting (ALL) [Re: PsilocyBen17]
    #22482222 - 11/05/15 06:01 PM (8 years, 2 months ago)

by "monoculture" you mean strain or isolate right?


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Offlinemushpunx
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Re: Agar isolation and fruiting (ALL) [Re: PsilocyBen17]
    #22482233 - 11/05/15 06:04 PM (8 years, 2 months ago)

Quote:

PsilocyBen17 said:
by "monoculture" you mean strain or isolate right?




Usually it refers to an isolated culture, no longer showing any sectors


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

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OfflinePsilocyBen17
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Re: Agar isolation and fruiting (ALL) [Re: mushpunx]
    #22482243 - 11/05/15 06:06 PM (8 years, 2 months ago)

gotcha


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OfflineMachiavelliavore
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Re: Agar isolation and fruiting (ALL) [Re: Erik77johansson77]
    #22482253 - 11/05/15 06:07 PM (8 years, 2 months ago)

Couple methods I've seen that look good:

Any sort of invitro grow, brf, straight grains, or some kind of other bulk sub.  Check out the violet invitro thread.  Grains are peferable since inoculating BRF with agar sucks.

Only issue with muda bottles is that you wouldn't really want to make LI for 8+ cultures.  You can try agar, but it will still be slow.  You can inoculate with grains, but at that point you might as well just spawn them.

Another method that I use is to a2g a single quart of spawn, then spawn it to two quarts of coir in a $1 foil lasagnia pan with a plastic lid.  Shotgun the lid with holes every 2.5" with a soldering iron, and use a piece of trash bag for liner.  Mist 1-2x a day, works very well.  Due to the high FAE of these things, use a casing.  Dollartree has them cheap, you can also pick'm up at target/walmart/etc... for a bit more.  At that point you might as well grab a 15qt sterilite and make a 3qt sub monotub.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Offlinemushpunx
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Re: Agar isolation and fruiting (ALL) [Re: Machiavelliavore]
    #22482354 - 11/05/15 06:31 PM (8 years, 2 months ago)

To inoculate BRF with agar you don't have to drop a wedge and use GE lid, you can sterilize a syringe of water, squirt some water onto your culture and then agitate with the flame sterilized needle and suck the myc water up for a fast LI
I


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

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AMU Q&A


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InvisibleSupalemonhaze
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Re: Agar isolation and fruiting (ALL) [Re: mushpunx]
    #22483179 - 11/05/15 09:11 PM (8 years, 2 months ago)

Quote:

mushpunx said:


This is why you don't want to just end up with one isolate from a culture.

You should keep transferring until you see sectoring (like in pic) and then take a transfer from *each* sector and then keep making transfers on each untill you have a whole bunch of isolates.

You should aim to end up with at Least 10 isolates from a culture to test out. If you just have one you have nothibv to compare it to, and unless your lucky it might be garbage. Usually 1 out of 10 is a keeper




Yea at first I just got so blinded with wanting an isolate that I just aimed on getting one. Then I saw RR's vids and realised how much of a fuck up im making. Lucklily I still had all my plates and the transfers I had were still sectoring it was easy to get back on the right track


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