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InvisibleTien
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Need help with strain isolation
    #4375410 - 07/06/05 01:09 PM (11 years, 5 months ago)

So I got my agar and stuff last week and made some plates.
I inoculated three petri-dishes with MEA using prints.
The strains are;

Pink Buffalo
Texas
PESA

The problem is that about 97% of the mycellium is all fluffy, I am bearly seeing any rhizos at all. So what should I do at this point? Try and isolate the tiny rhizos? The Texas culture is not showing any rhizmorphic growth at all, which is very disappointing..I had high hopes for this strain. What should I do with this plate if it doesn't grow any rhizo mycellium?


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Invisibletahoe
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Re: Need help with strain isolation [Re: Tien]
    #4375737 - 07/06/05 02:50 PM (11 years, 5 months ago)

are the petris fully colonized? alot of times rhizos dont form on the first plate so you could transfer any spot. But if you want then the rhizos should shoot out past the cotton and use thm for the transfer. You could use the little rhizos that you have


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Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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Offlinescatmanrav
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Re: Need help with strain isolation [Re: tahoe]
    #4375780 - 07/06/05 03:05 PM (11 years, 5 months ago)

I'd go for a section with as much rhizo on it as you can. The next transfer should have more rhizo on it after that and so on and so on.


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"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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Offlinefreakaz0id
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Re: Need help with strain isolation [Re: tahoe]
    #4375783 - 07/06/05 03:06 PM (11 years, 5 months ago)

why not just inoculate with the dishes you got... and pick a nice shroom to clone?
i guess there is no better isolation...?


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InvisibleTien
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Re: Need help with strain isolation [Re: freakaz0id]
    #4375816 - 07/06/05 03:18 PM (11 years, 5 months ago)

Thanks for the input guys, I'm going to isolate the rhiziest spots I can.

Quote:

freakaz0id said:
why not just inoculate with the dishes you got... and pick a nice shroom to clone?
i guess there is no better isolation...?




I want to isolate a strain first so I can have quick colonization, rhizo mycellium, etc.

If I was to inoculate my substrate with the multi-spore cultures that I have, it would be a guess what kind of mycellium is going to dominate.

Pluto


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OfflineRogerRabbitM
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Re: Need help with strain isolation [Re: Tien] * 1
    #4377475 - 07/06/05 10:17 PM (11 years, 5 months ago)

It's very normal not to see the rhizomorphic growth until you make few transfers. With multispore inoculation on agar, you have hundreds if not thousands of strains growing together and all over on top of each other. Remember, any two spores that germinate and 'pair up' by forming a clamp connection is the very definition of a 'strain'. Just dig in and take a very small piece of mycelium no larger than a grain of rice. Move this to a new dish.

When this dish begins to grow out, you may or may not begin to see sectors. It might take a second transfer before you get few enough strains to see the individual sectors. Take the best looking growth again and move it to a fresh petri dish. Eventually, when you hold a dish up the the light and look from the back(agar) side, you will see individual sectors like the spokes on a wheel.

In the picture below, the dish on the left is the result of two transfers from the original spore swipe. Prior to the second transfer, the rhizomorphic mycelium was buried in the mass of strains and couldn't be differentiated, just like yours are. The rhizomorphic sections are now clearly showing up at ten o'clock and two o'clock. Each of those sections has four or five individual sectors that will be isolated out next time I do agar work in a day or two. The remainder of the dish will be discarded. The dish on the right is a single sector isolate, as you can see the entire dish is rhizomorphic and when held up to the light, there is no further sectoring. I fruit out each individual sector to determine the best fruiting isolate. Many isolates are mediocre fruiters, but if you want to find the one that will deliver those monster flushes time and time again, this is how it's done. I keep a small piece from each petri dish in the refrigerator until I've found the best fruiting isolate, then I get that piece out of the fridge and grow it out. I always keep a piece from the original petri dish in test tubes in the refrigerator so I can go back to that strain for years to come with no fear of senescense. It's a lot of work, but hey. . .It's a hobby right?
RR


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



semper in excretia sumus solim profundum variat


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InvisibleTien
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Re: Need help with strain isolation [Re: RogerRabbit]
    #4377763 - 07/06/05 11:45 PM (11 years, 5 months ago)

That cleared up a lot, really. This is the info I have been looking for all along. Thanks a bunch RR, 5 shrooms for you brother.

Pluto


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OfflineNo_Life_G33k
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Re: Need help with strain isolation [Re: Tien]
    #4451612 - 07/25/05 06:41 PM (11 years, 4 months ago)

Look for the link in RR's post that points to his other thread... the pix explain a lot.....

http://www.shroomery.org/forums/showflat...rue#Post3991641


Mega props to Roger!!


--------------------
Hot Water Pasteuriztion - A How To Pictorial
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4527808/an/0/page/0

Make your own shroom chocolates
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4500682/an/0/page/5

guide with pix.


Make Your Own Magnetic Stirrer! How-To Tek w/ pix
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5865949/an/0/page/3


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Re: Need help with strain isolation [Re: No_Life_G33k]
    #19660555 - 03/06/14 09:04 PM (2 years, 8 months ago)

how many transfers is too many?


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OfflineRogerRabbitM
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Re: Need help with strain isolation [Re: eelrd72]
    #19660770 - 03/06/14 10:15 PM (2 years, 8 months ago)

I doubt he's still looking after 8 years.  Don't bump old threads without something important to add.
RR


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



semper in excretia sumus solim profundum variat


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Re: Need help with strain isolation [Re: RogerRabbit]
    #19662209 - 03/07/14 07:44 AM (2 years, 8 months ago)

Can't ask a question on an old thread? without bumping it?  oh sorry I just bumped it again. I asked the question because I wanted to know how many transfers are too many and I also wanted to save this post for future reference. Is there a way to save post without bumping them?


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OfflinePussyFartM
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Re: Need help with strain isolation [Re: eelrd72]
    #19662223 - 03/07/14 07:52 AM (2 years, 8 months ago)

Quote:

eelrd72 said:
Is there a way to save post without bumping them?



Favorites or Bookmarks in your browser.


--------------------
                 
THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT!
A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD,
SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List,
EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE
  ^*~.,,.~*^*~.,_,.~*^*~.,_SHIT FOR NOOBS_,.~*^*~.,_,.~*^*~.,_,.~* 


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OfflineRogerRabbitM
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Re: Need help with strain isolation [Re: PussyFart]
    #19662643 - 03/07/14 11:38 AM (2 years, 8 months ago)

No amount of transfers is too many.  Isolate to single sectors and test fruit them all if you want to find that super strain.
RR


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



semper in excretia sumus solim profundum variat


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Offlineeelrd72
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Re: Need help with strain isolation [Re: RogerRabbit]
    #19735682 - 03/23/14 10:36 AM (2 years, 8 months ago)

when you purchase a print...from sponsor..are those prints from isolates?


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OfflineStromrider
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Re: Need help with strain isolation [Re: eelrd72]
    #19735958 - 03/23/14 12:07 PM (2 years, 8 months ago)

Quote:

eelrd72 said:
when you purchase a print...from sponsor..are those prints from isolates?




Doesn't matter


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Offlineeelrd72
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Re: Need help with strain isolation [Re: Stromrider]
    #19736158 - 03/23/14 01:02 PM (2 years, 8 months ago)

is that because..when i take the print to agar....i make a multispore slide again?


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OfflinePussyFartM
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Re: Need help with strain isolation [Re: eelrd72]
    #19736211 - 03/23/14 01:20 PM (2 years, 8 months ago)

Quote:

eelrd72 said:
is that because..when i take the print to agar....i make a multispore slide again?



Spores are starting over from scratch, with unknown genetics.

It does not matter if the spores came from a clone, an isolate, or MS.


--------------------
                 
THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT!
A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD,
SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List,
EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE
  ^*~.,,.~*^*~.,_,.~*^*~.,_SHIT FOR NOOBS_,.~*^*~.,_,.~*^*~.,_,.~* 


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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