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Offlineeltrkbrd
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Registered: 02/23/07
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Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis?
    #6808753 - 04/19/07 01:12 PM (16 years, 11 months ago)

I have a 250ml Eberbach Container and Waring Blender Base.

My goal is to store the solution longterm in 24ml autoclavable dram vials with airtight caps using distilled water + the blended mycelium that was scraped off the petris.

I have six 3 section petris fully colonized with a pure culture (B+) after completing 1 transfer. The mycelium has already started creeping up the walls so I have to do something with it now before it's too late.

All the petris were grown out using the inside stipe of a single fruit body (B+), can I just combine/blend the mycelium from all 6 petris or is it smarter to blend them up separately?

Since this is a stipe cloned to Agar, does anastomosis leave even the remote chance that I have different strains on these plates?

It would be a lot easier to blend them all in one shot.

-TIA


--------------------
My way of giving back- Thanks Shroomery...
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OfflineRogerRabbitM
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6808809 - 04/19/07 01:25 PM (16 years, 11 months ago)

I wouldn't mix them. If one has a contaminant spore laying dormant, you'll fubar the whole batch by blending them together. You also don't want to store anything with air tight lids. The mycelium will suffocate and die, even under refrigeration.
RR


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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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InvisiblePsychoslut
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: RogerRabbit]
    #6808844 - 04/19/07 01:36 PM (16 years, 11 months ago)

so when you store a culture in a test tube it is not air tight?


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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InvisibleRoadkillM
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6808852 - 04/19/07 01:38 PM (16 years, 11 months ago)

Long Term Storage of Mycelium in Distilled Water...

does work.


tc


--------------------
Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!


Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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Offlineeltrkbrd
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: RogerRabbit]
    #6808905 - 04/19/07 01:48 PM (16 years, 11 months ago)

Hi RR,

Thanks for the advice! What if I leave the cap on loosely and wrap parafilm around it?

The challenge with the method you suggest for blending each dish separately is going to be my sterile technique.

I have 7 dishes which is enough to make at least 7 vials, so what if I sterilize each vile 3/4 full with distilled water and then when I go to transfer the mycelium in to the eberbach container I wipe it down and set it into the glovebox and after sitting (for air to still), pop the lid on the eberbach container, transfer the culture into the container and reseal the lid, remove from the glovebox and blend.

Then wipe down eberbach container, place back in glovebox, pour sterile distilled water from vial into Eberbach, transfer mycelium from dish, remove from glovebox and blend. Repeating the process with the remaining dishes and vials.

Will this work, or is there a better method for long term storage of these cultures?

I did get a good deal on the eberbach, so I figured it would come in handy for this, right?

Also, from the Shroomery Agar guide on "Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures": http://www.shroomery.org/8509/Resurrecting-a-Better-Method-for-Long-Term-Storage-of-Mushroom-Cultures

<quote>"Screw the lid on tight and wrap Parafilm around the top of the vial to make sure it is airtight."</quote>

Thanks RR!

P.S.
I'm excited to hear the news update today on your upcoming DVD release.


--------------------
My way of giving back- Thanks Shroomery...
A For Newbs by Newbs Cultivation Resource Journal: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6748262/an/0/page/0

Edited by eltrkbrd (04/19/07 01:55 PM)

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Offlineeltrkbrd
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6808967 - 04/19/07 02:03 PM (16 years, 11 months ago)

Hi Road!

When you say "Long Term Storage of Mycelium in Distilled Water...does work."

Is this referring to the document exactly as described in the Agar section here on the Shroomery?

-Thanks


--------------------
My way of giving back- Thanks Shroomery...
A For Newbs by Newbs Cultivation Resource Journal: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6748262/an/0/page/0

Edited by eltrkbrd (04/19/07 02:04 PM)

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InvisibleRoadkillM
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6808977 - 04/19/07 02:05 PM (16 years, 11 months ago)

no

I was taught a method of doing so...

by an old mycologist.

:smile:


--------------------
Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!


Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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InvisiblePsychoslut
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: Roadkill]
    #6808990 - 04/19/07 02:07 PM (16 years, 11 months ago)

i miss pappy:(


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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InvisibleRoadkillM
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Registered: 12/11/01
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: Psychoslut]
    #6809007 - 04/19/07 02:11 PM (16 years, 11 months ago)

Quote:

Psychoslut said:

i miss pappy:(




I know...

alot of people do!~


he pops in and visits in the Mod forum now and then.

:smile:


--------------------
Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!


Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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Offlineeltrkbrd
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Registered: 02/23/07
Posts: 65
Last seen: 1 year, 10 months
Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: Roadkill]
    #6809083 - 04/19/07 02:36 PM (16 years, 11 months ago)

So you're using a different method? :smile:

Any advice for this noob in regards? :confused:


--------------------
My way of giving back- Thanks Shroomery...
A For Newbs by Newbs Cultivation Resource Journal: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6748262/an/0/page/0

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Offlineboatshow
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Registered: 09/01/05
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6809169 - 04/19/07 03:01 PM (16 years, 11 months ago)

Special mod forum? No fair. I bet that's where all the good stuff is.


--------------------
I saw the best minds of my generation destroyed by madness, starving hysterical naked,
Dragging themselves through the negro streets at dawn looking for an angry fix,
Angelheaded hipsters burning for the ancient heavenly connection to the starry dynamo in the machinery of night

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Offlineeltrkbrd
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Registered: 02/23/07
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: boatshow]
    #6809193 - 04/19/07 03:09 PM (16 years, 11 months ago)

Yea, where's the tiki lounge?


--------------------
My way of giving back- Thanks Shroomery...
A For Newbs by Newbs Cultivation Resource Journal: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6748262/an/0/page/0

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Offlineeltrkbrd
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Last seen: 1 year, 10 months
Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eltrkbrd]
    #6809550 - 04/19/07 04:59 PM (16 years, 11 months ago)

Anyone have an answer for me regarding the technique?

I'm a bit confused about the Airtight cap being sealed vs. loose/wrapped with parafilm and the sterile technique I need to employ here for blending multiple times going in and out of the glovebox.

-TIA


--------------------
My way of giving back- Thanks Shroomery...
A For Newbs by Newbs Cultivation Resource Journal: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6748262/an/0/page/0

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OfflineeVenom
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: Roadkill]
    #7391079 - 09/10/07 08:38 AM (16 years, 6 months ago)

Quote:

Roadkill said:
no

I was taught a method of doing so...

by an old mycologist.

:smile:




I am really interested in this method could you describe it!?

I don't find it anywhere!!


--------------------
eVenom

"Some of us like a nice buzz, while others want to climb a screaming volcano naked to meet god."
RR

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InvisibleRoadkillM
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: eVenom] * 1
    #7391177 - 09/10/07 09:20 AM (16 years, 6 months ago)

You really need a flowhood, or a glove box, or a still air box to do this procedure.

everything must be kept extremely sterile.



pressure cook some distilled or purified water in a Pint or Quart jar...
let that cool down to room temp.
then put that in front of your flowhood or inside your glovebox or still air box.

clean the top of your colonized petri dish with Isopropyl Alcohol..
then put your petri dish in front of the flowhood or inside your glovebox or still air box.

Put some sterile test tubes in front of the flowhood or inside your glovebox or still air box.

then put your sterilized tools such as a Scalpel, Exacto knife, Inoculation Loop, Denist Tools,
what ever you use...in front of the flowhood or inside your glovebox or still air box.



Open up the petri dish with the colonized mycelium...
scrape the mycelium off the agar trying to just get the mycelium without any agar...
with whatever tool you choose or prefer.
put the mycelium into the test tube.
then add some sterilized water from the jar you pressure cooked.
put the top onto the test tube.
rap the test tube with several layers of paper towels.
put into a ziplock freezer bag.

store in the frig.

wala!~

:smile:



tc


--------------------
Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!


Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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OfflineeVenom
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Re: Eberbach for Long Term Storage of Mycelium in Distilled Water - Anastomosis? [Re: Roadkill]
    #7391561 - 09/10/07 11:22 AM (16 years, 6 months ago)

awesome!!!!

Thanks a lot!!!!


--------------------
eVenom

"Some of us like a nice buzz, while others want to climb a screaming volcano naked to meet god."
RR

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