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InvisibleJoshua
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Wild Strain Isolation - Print to Print **complete**(1/16)
    #1238096 - 01/21/03 10:35 PM (14 years, 5 months ago)

This log will document the steps necessary to isolate a wild strain from print to print.

I obtained a wild print in a trade. The print was taken from a non-sterile fruit.

The following are steps in one particular procedure. Many modifications can be made to get to the same endpoint.

The first step in isolation is to streak a plate with the spores. This method consists of taking an inoculation loop and transfering spores from the print to the surface of an agar plate. I used I scalpel for this step and an agar plate with an antibiotic mixed in. I tend to inoculate the plates at points rather than streaking. Streaking requires pulling the inoculation loop across the surface in a sweeping motion. The agar used was Malt Extract Agar w/ .05 g gentamycin/ liter. The gentamycin prevents bacterial growth. I use gentamycin because is survives higher temperatures, other antibiotics tend to break down at the temps reached in my PC.

This is what the plate looked like after 5 days according to my records (which are probably faulty as this is much more growth than should be expected after 5 days.) This growth is very nice considering it came from a wild print.



I will give out more info regarding the origin of the strain after I talk to the user that provided the strain to me. I will be adding steps as frequently as time allows. Currently this strain is much further along than I have shown here. It is now colonizing millet.

Joshua


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"Life After Death is Saprophytic!"


Edited by Joshua (01/16/04 02:03 PM)


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InvisibleG a n j a
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1238133 - 01/21/03 11:38 PM (14 years, 5 months ago)

Thanks for this joshua this will be intereesting to follow :smile:


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OfflinestonErollEr1
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1238238 - 01/22/03 02:35 AM (14 years, 5 months ago)

Nice!
I will follow this thread...closely.
Is it what i think it is? :wink:

peace...


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InvisibleJoshua
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Re: Wild Strain Isolation - Print to Print [Re: stonErollEr1]
    #1238531 - 01/22/03 05:46 AM (14 years, 5 months ago)

Yes it is. Please keep the info to yourself until I announce the specifics. I'd like to talk to the provider of the print regarding the naming of this strain.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


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OfflinestonErollEr1
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1239005 - 01/22/03 09:18 AM (14 years, 5 months ago)

Sure will...bro!

peace..


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OfflineSeril
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1239417 - 01/22/03 11:44 AM (14 years, 5 months ago)

exciting! :grin:


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Seril,
Political language -true of all political parties- is designed to make lies sound truthful and murder respectable, and to give solidity to pure wind..


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Invisibleangryshroom
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1239469 - 01/22/03 11:57 AM (14 years, 5 months ago)

Yes, very cool indeed :smile:


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OfflineDymentia
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1240226 - 01/22/03 04:04 PM (14 years, 5 months ago)

what is this all about?
Quote:


Is it what i think it is? :wink:




Quote:

Yes it is. Please keep the info to yourself until I announce the specifics.




the suspense is killing me, i must know! :smile:
 


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Offlinedebianlinux
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Re: Wild Strain Isolation - Print to Print [Re: Dymentia]
    #1240313 - 01/22/03 04:37 PM (14 years, 5 months ago)

ok, i'm hooked, it's obviously believed to be a new strain.


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Invisibleblahblahblah
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1240663 - 01/22/03 06:48 PM (14 years, 5 months ago)

Quote:

I will give out more info regarding the origin of the strain after I talk to the user that provided the strain to me. I will be adding steps as frequently as time allows. Currently this strain is much further along than I have shown here. It is now colonizing millet. 



I hope you have some more pictures of the petris and such, THAT is what I want to see  :grin:


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InvisibleJoshua
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Re: Wild Strain Isolation - Print to Print [Re: blahblahblah]
    #1242178 - 01/23/03 08:39 AM (14 years, 5 months ago)

To isolate a strain is also called sectoring. After the original inoculations grew out I chose areas of healthy/vigorous growth and transfered squares of the healthy sections to a new plate. These are pics of the first transfer grown on Dog Food agar.

I think this one is smiling at all of you Shroomerites!



It is always best to work with multiple plates, as some may succumb to contamination. It also increases your chances of finding a healthy strain as some isolates will perform better than others.

The branching you see is an indication of healthy growth. This type of growth is called rhizomorphic growth. Rhizomorphic growth grows faster and forms hyphal knots more readily than cottony growth. The rhizomorphic growth is good and is what I am looking for. I would pick the best growth off of these plates and transfer them to new plates.



Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


Edited by Joshua (01/23/03 08:45 AM)


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InvisibleG a n j a
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1242436 - 01/23/03 10:17 AM (14 years, 5 months ago)

I'm in the process of trying this for the first time so thank you for this.
Very clear pictures.How long did it take for the first plate to fill up like that and at what temp?

Cheers


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Invisible@cro
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1242438 - 01/23/03 10:17 AM (14 years, 5 months ago)

Those are some nice lookin rhizos there, if this is going to be a new strain I can see it quickly becoming popular.  Good Work, have fun.  :smile: 


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InvisibleJoshua
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Re: Wild Strain Isolation - Print to Print [Re: G a n j a]
    #1243747 - 01/23/03 06:10 PM (14 years, 5 months ago)

The growth you see in those plates is probably 5 days old.

Here is a petri of a different isolation I am working on as well. I am showing this one to show the difference of what a good isolation and a not-so-good isolation may look like.

This was the growth from the initial inoculation.



This is the growth after 2 transfers.


I hope you can see the difference in the rhizomorphic growth between the two isolates.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


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InvisibleG a n j a
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1244149 - 01/23/03 08:52 PM (14 years, 5 months ago)

Yep big difference
i can also almost read what you blacked out in picture 1 lol :tongue:
but only almost :frown:    lol


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InvisibleAlkaloids
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1244204 - 01/23/03 09:20 PM (14 years, 5 months ago)

Good work as usual Joshua. I'll be following this as I am going to post something similar in the near future.
What are those container you are using? The agar tranfers look unusually thick.


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InvisibleJoshua
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Re: Wild Strain Isolation - Print to Print [Re: Alkaloids]
    #1244216 - 01/23/03 09:24 PM (14 years, 5 months ago)

I have polypropylene and Pyrex dishes. The polypropylene are quite a bit deeper than regular petris. I like to pour my plates thicker than most. Perhaps w/ more experience I will pour them thinner and conserve on supplies.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


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OfflineShrewDigsby
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Re: Wild Strain Isolation - Print to Print [Re: G a n j a]
    #1244238 - 01/23/03 09:33 PM (14 years, 5 months ago)

This is an awesome log...just like all the rest :laugh:

I want to learn, I want to learn :grin:

Ganja, you have great logs too, you should post your latest. 


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Marijuana is a horticultural plant.  Hemp is an industrial weed.  I believe they were both provided to us by GOD to use and enjoy.


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OfflinestonErollEr1
The Psilocybinsolution
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Re: Wild Strain Isolation - Print to Print [Re: ShrewDigsby]
    #1244511 - 01/24/03 12:48 AM (14 years, 5 months ago)

Looking nice Joshua.
Very healthy rhizos there.

Peace..


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InvisibleZen Peddler
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Re: Wild Strain Isolation - Print to Print [Re: Joshua]
    #1244542 - 01/24/03 01:24 AM (14 years, 5 months ago)

I think some points should be raised here.
Firstly Antibiotics such as gentamycin dont actually stop bacterial contaminants but they can limit their prevelance. Bacterial as opposed to molds which are more of a problem, appear as liquidy spots on the plate. Because you are streaking the spores across the plate you should be able to quickly get some germination points that dont display any bacteria which you can simply transfer to h202 agar.
Antibiotics will not effect the germination of contaminants like everyone's favuorite green molds (Pencillium etc.) and as these have spores, they can quickly spread and contaminate an entire plate.
As Joshua has stated, that is why it is important to innoculate five or six plates and quickly transfer to h202 agar to kill contaminant spores and clean up the culture. From there you can isolate rhizomorphic sectors.
The last point i wanted to make is that rhizomorphic mycelia does not guarantee strong fruiting performance. With cubensis once the plate is fully colonised and exposed to light, it will fruit. Even if you isolate some sectors that seem linear or even cottony, they may perform better than the rhizomorphic mycelia that Stamets loves so much - there is a post regarding this issue in advanced forums right now. So test every isolate - nice looking mycelia or otherwise. Good practise as most mushrooms wont fruit off agar - youll never be able to test this with Hy.Capnoides or Lepista Nuda.
BTW - most cubensis prints ive received that ive streaked on agar have contaminated worse than wild prints. Due to the stagnent nature of fruiting chambers im assuming that there are very, very few cubensis that are grown in sterile environments. My point being that if you can grow out normal cubensis on agar, you can grow out wild varieties just as easily.
This goes for pan cyans as well, they are pretty easy to domesticate to fruit well - easier than the weird Ps.Eucalypta and Ps.Stuntzi ive been working with.


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Edited by Zen Peddler (01/24/03 01:27 AM)


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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation >> Mushroom Cultivation Archive >> Grow Logs

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