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Amazon Shop for: ½ Pint Jars, 10cc Syringe, Autoclave, Laminar Flow Hood, Pressure Cooker

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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
*NEW* Alien Broth Tek --- with pictures!
    #1656554 - 06/23/03 08:10 PM (13 years, 5 months ago)

Alien Broth Tek



Hello fellow travelers
in space. I feel I should share a wonderful story:


I have a friend that was
abducted by aliens and was given the following instructions and photographs. It
appears to be instructions for the propagation and archival of live fungal
mycelium. Much of it taken directly from the industrial processes of this
strange alien race. I suggest that this process be used for research purposes
only and applies to many fungal species, many of which are inedible.


Now that we have dispensed
the formalities:


Basic Recipe:
1.50
Liters of Spring Water (for mineral content)
35 grams Maltose
Monohydrate
32 grams Dextrose (anhydrous glucose)

3 grams Thiamine Hydrochloride (vitamin B1)



Required Items:

36 Septa Vials (40ml. - there are 72 in a standard "field pack")
Sterile 3cc. Syringes with luer-lock tip

22 gauge 1.5" Sterile Hypodermic Needles (luer-lock)


Possible Substitutions:

High Maltose Refined Rice Syrup Solids in place of Maltose and Dextrose
(available from your local home brew supply) and 1cc. Insulin syringes for 3.cc
luer-lock syringes.



Instructions:







Weigh and mix maltose, glucose and thiamine with spring water. Using a sterile
60cc. syringe add 30 ml. of broth solution to your 40 ml. septa vials.










Cook these in a pressure cooker at 15 P.S.I. for 20-25
minutes.







Allow to cool to room temperature an decompress
slowly to avoid boil over. Remove lid and seal vials tightly and immediately.
Now here is the tricky part. The aliens did a sterile loop from active spores in
sterile alien broth solution to get my initial mycelium. Technically any live
mycelium added to broth solution should work once it has been established that
the mycelium is live and free of other organisms.  Once these vials had
nothing but mycelium growing in them, they shook them up and distributed 0.1cc.
(10 units) to each of my autoclaved and sealed 40 ml. septa vials. Visible
growth appears in 2-3 weeks with a good "snot wad" forming in 1-2 months.










Once growth is established in the septa vials use PF,
Hawk or other growth medium method.  Shake vials and inject with a long
needle 1cc. of solution into your media to initiate rhyzomorphic mycelium growth
(or dump it in a sprayer, shake it up and visit your local cow fields to spray
new cow patties).








Other Information:


The aliens have tried many broth combinations,
including a straight glucose to make both 5% and 10% solutions. They found the
above formula to be the most prolific. Although the mycelium does grow in
nothing but glucose and water, the growth was slow and small and barely at all
in the 10% solution. If you use rice syrup solids make a 5% solution (50 grams
per liter of water).





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InvisibleHippie3
mycotopiate
 User Gallery

Registered: 11/07/99
Posts: 3,090
Loc: mycotopia.net
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656812 - 06/23/03 09:58 PM (13 years, 5 months ago)

where are you geting your vials ?


--------------------
Admin @ mycotopia.net
Mycotopia


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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Hippie3]
    #1656820 - 06/23/03 10:02 PM (13 years, 5 months ago)

A standard google search for Septa Vial 40 should reveal many sources. Also known as an EPA Field Sample Vial.


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InvisibleAinasko
Oksania
Registered: 04/26/03
Posts: 694
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656847 - 06/23/03 10:17 PM (13 years, 5 months ago)

Here's a source for the vials.

They aren't very cheap, but they do serve a useful purpose. I use them for condensed storage of spore solutions.

I just draw half a cc of condensed spore solution per 10 cc's of sterile water.

It makes preparing syringes clean and simple.





--------------------
Ainasko is my name backwards. I'm a girl!


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InvisibleHippie3
mycotopiate
 User Gallery

Registered: 11/07/99
Posts: 3,090
Loc: mycotopia.net
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656848 - 06/23/03 10:17 PM (13 years, 5 months ago)

lol
that's not what i asked, but ok.
:ooo:


--------------------
Admin @ mycotopia.net
Mycotopia


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InvisibleHippie3
mycotopiate
 User Gallery

Registered: 11/07/99
Posts: 3,090
Loc: mycotopia.net
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Ainasko]
    #1656851 - 06/23/03 10:19 PM (13 years, 5 months ago)

thx for the link,
you weren't kidding about them being expensive.
think i'll just stick with my tyvek-filtered jars for now.


--------------------
Admin @ mycotopia.net
Mycotopia


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InvisibleAinasko
Oksania
Registered: 04/26/03
Posts: 694
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Hippie3]
    #1656855 - 06/23/03 10:22 PM (13 years, 5 months ago)

If you live in a larger city, you might also want to try looking in your local phone book for laboratory suppliers. If you find a friendly pharmacist, it wouldn't hurt to ask him too.


--------------------
Ainasko is my name backwards. I'm a girl!


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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Hippie3]
    #1656856 - 06/23/03 10:22 PM (13 years, 5 months ago)



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



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InvisibleOscuro_lobo
Dark Wolf

Registered: 04/15/02
Posts: 813
Loc: it doesn't really matter.
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Ainasko]
    #1656867 - 06/23/03 10:27 PM (13 years, 5 months ago)

Those vials look like the same kind of bottles insulin comes in, with the foil tops that are impossibl to remove without cutting them.

How do you get those caps sealed on, after sterilizing the water in them, and getting spores into them and such.

I would imagine, with such a small opening at the top it would be very difficult to do this, and maintain sterility without a glovebox/flowhood.

Do you just make a sporesyringe the normal way, and inject it into the sterile bottles? Wouldn't that pressurize them?

Also, how do you make such a concentrated spore solution? Like, how do you keep it consistent, so that your measurements work out.

Like.. 3 medium prints to 10cc's water?

-ol


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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656883 - 06/23/03 10:31 PM (13 years, 5 months ago)

The nice thing about the EPA Septa Vials is that they can be autoclaved over and over, and replacements can be bought in bulk for worn out teflon seal/injection sites. The use of high gauge needles insures long life for the teflon heads. A good standard field gauge is 22.


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



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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656886 - 06/23/03 10:32 PM (13 years, 5 months ago)

The tops screw off and seal tight. They are very nice to work with.


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



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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656895 - 06/23/03 10:35 PM (13 years, 5 months ago)

Pressure is OK. Hence the reason for autoclaving with the caps loose, and cooling to room temp. to get normal atmospheric pressure. Vacutainers would work fine. Although mycelium will grow in an anaerobic environment, they grow faster in the presence of regular atmosphere. They have given strong consideration to compressing the vials with a nitrox diving gas mixture, or even pure 02 (careful!) if you have access to medical or welding gasses. Once autoclaved a regular spore syringe can be used to start the master vial(s) that would be used to distribute live mycelium to other archive vials. Yet to discover how long mycelium will live in these vials placed in the fridge or safety deposit box, but 3 years have been proven by the aliens, and feasably indefinately.


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



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InvisibleAinasko
Oksania
Registered: 04/26/03
Posts: 694
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Oscuro_lobo]
    #1656913 - 06/23/03 10:41 PM (13 years, 5 months ago)

I am a student in a biological science lab. I have the unfair advantage of of air showers, flow hoods and autoclaves in place of pressure cookers.

You can purchase the vials with screw caps, the caps pre-crimped or you can purchase the vials and caps separately. If you buy them separately, you'll also need to buy a crimper to join them.

If you buy them pre-sealed and attached, they are no longer easily autoclavable and you'll have to trust in the supplier.

Do you just make a spore syringe the normal way, and inject it into the sterile bottles? Wouldn't that pressurize them?

When I do use pre-sealed vials, a trick I learned is to first inject the sealed vial with an empty syringe and draw the plunger to create a vacuum. When I inject again to fill the vial, pressure becomes neutral .

We maintain spore consistancy with a spore density test, but it's not really necessary. Just a few large, dark prints will produce a highly concetrated solution that can be diluted for later use.


--------------------
Ainasko is my name backwards. I'm a girl!


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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Ainasko]
    #1656938 - 06/23/03 10:48 PM (13 years, 5 months ago)

See the reply above yours. As a student you might have access to educational/industrial mycology methods and research that might prove very enlightening.


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



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InvisibleAinasko
Oksania
Registered: 04/26/03
Posts: 694
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1656948 - 06/23/03 10:51 PM (13 years, 5 months ago)

Quote:

metaophion said:
See the reply above yours. As a student you might have access to educational/industrial mycology methods and research that might prove very enlightening.




Not really. The only advantage I have is equipment and a sterile environment.

I can be taught the latest cultivation techniques, but I can never be taught experience.


--------------------
Ainasko is my name backwards. I'm a girl!


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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: Ainasko]
    #1657970 - 06/24/03 08:55 AM (13 years, 5 months ago)

That is quite an advantage!
One thing about the Septa Vials is once you work with teflon/silicone heads - you will never go back to rubber heads, which tend to wear out after repeated autoclavings and punctures... They think a box of 72 40ml. vials with screw on caps should be about $40.00USD plus the septums on the screw-tops are replaceable.


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



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Offlinemetaophion
stranger thanstrange
Registered: 06/23/03
Posts: 28
Loc: Seminyak, Bali - Indonesi...
Last seen: 12 years, 7 months
Re: *NEW* Alien Broth Tek --- with pictures! [Re: metaophion]
    #1658500 - 06/24/03 02:46 PM (13 years, 5 months ago)



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Amazon Shop for: ½ Pint Jars, 10cc Syringe, Autoclave, Laminar Flow Hood, Pressure Cooker

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