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InvisibleSpookerShroom
Shroomerite
Registered: 06/12/06
Posts: 244
Creamcorn cloning in a vacutainer?
    #5944101 - 08/08/06 01:04 PM (17 years, 5 months ago)

Ive been doing some cloning following CC's stabbing (coring) tek and its been working great for me...

Now I want to start storing some long term master cultures in the fridge, so I got a few 10cc vacutainers.

My question is instead of taking some LC from a jar that was made from the core.....Could I just fill the syringe with Karo solution and sterilize (instead of water in CC's tek), and the take my core of my selected fruit, and inject the solution/core right into the vacutainer, letting it grow out, then place in the fridge?

Main reason is just because I want to skip the step of aspirating some LC from a jar and injecting into the 'tainer.....The only problem I can see with putting the core directly into the long term storage 'tainer is the core itself....will it last w/o rotting as long as the LC does under refridge?

Thanks guys/girls


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InvisibleTippinthru
contented

Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
Re: Creamcorn cloning in a vacutainer? [Re: SpookerShroom]
    #5944180 - 08/08/06 01:27 PM (17 years, 5 months ago)

For long term tissue storage.

A needle biopsy tissue sample should be stored in sterile water.

There should be no air/gas, or nutrients in the water.

That circumstance places the tissue in stasis.

It there is air, and nutrients, the tissue does not go into stasis.

It tries to grow.

In doing so, it's viability will decrease, over time.

Vacutainers work great for long term tissue storage.

At a later date, if you are worried about extracting the tissue sample.

Simply use a larger bore needle, to do so.


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Perfection is attained by slow degrees; it requires the hand of time...
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InvisibleSpookerShroom
Shroomerite
Registered: 06/12/06
Posts: 244
Re: Creamcorn cloning in a vacutainer? [Re: Tippinthru]
    #5944298 - 08/08/06 02:13 PM (17 years, 5 months ago)

Thanks tipp, always can count on you for a great response :smile:

So your saying I should just sterilize the syringe with just water (as in CC's tek), go ahead and take the core (biopsy), and then inject the core with its water into the vacutainer? And when I want to make more of that specific fruit, just go in with a big needle and suck up the core and use it to start a LC a la CC's tek?

I was wondering about how long a 10cc portion of karo LC would last under refrig....im guessing not as long as the biopsy stored in water im guessing....only thing is its not truly a master culture right? I was looking for something that I could take 1-2cc's from every month to start a new LC....with just the biopsy its a one time deal..unless I take 10 cores lol?

Sorry for the ramble. And thanks again, yout posts are a joy to read


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InvisibleTippinthru
contented

Registered: 04/07/05
Posts: 1,131
Loc: "The Garden"...
Re: Creamcorn cloning in a vacutainer? [Re: SpookerShroom]
    #5944475 - 08/08/06 03:16 PM (17 years, 5 months ago)

An LC will last 3 or 4 months, if carefully, and constantly refrigerated.

Remember also, any LC that is refrigerated, should be brought back to room temperatures, before use.

Another storage option is to take a large syringe, at least 20ml, or preferably larger.

Aspirate as much LC myc into it, as possible.

Replace the needle sheath, wrap syringe in a protective covering (foil, cling wrap, ect).

Then, stand the syringe in a refrigerator, with the needle pointing straight up for a couple days.

All the mycelium will settle to the bottom of the syringe.

Then, holding the syringe upright, very carefully squirt as much nutrient solution, out of the syringe, without squirting out the mycelium.

Once done, refill the syringe with sterile distilled water.

What that does is remove most of the nutrients.

With those removed, the mycelium will store much longer.

You can use a few ml, from that master, to start LC's, as you want.

You want to maintain that master syringe, as sanitary, as possible, at all times.


--------------------
Perfection is attained by slow degrees; it requires the hand of time...
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Offlinetrentallica
Lord Trentnar
Male User Gallery

Registered: 05/01/06
Posts: 797
Loc: virgina Flag
Last seen: 8 years, 8 months
Re: Creamcorn cloning in a vacutainer? [Re: Tippinthru]
    #5944729 - 08/08/06 04:45 PM (17 years, 5 months ago)

i believe this is the tek that tripp is talking about. you can store your prints, or clone coultures for like 10 years in plane water.

http://www.shroomery.org/index.php/par/23543



" Storage Method:

Obtain dram vials from your laboratory supplier and fill them about half full (about 3mL) with distilled water, loosly cap the vials and sterilize them in a pressure cooker for 30 minutes @250 F.

Half dram vials or test tubes with screw caps would also work well.

*** About six milliliters of sterile distilled water is pipetted aseptically into a freshly growing culture. The fragments of hyphae are dislodged by lightly scraping the aerial growth with the same pipette, and the resulting suspension is withdrawn and transferred to a sterile glass vial. Put plenty of inoculum into each vial to insure success.

Screw the lid on tight and wrap Parafilm around the top of the vial to make sure it is airtight.

When you come back in a few years,you would not want to find that the water had evaporated.

Store the vials at room temperature away from direct sunlight. A bookshelf or wall cabinet is an excellent place. If conditions deteriorate, and the room should become unbearably hot, the vials can be refrigerated, but that is not normally necessary.

In the distilled water envirnment, the mushroom culture enters a dormant state, and it is held in stasis. "


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The only ones for me are the mad ones..........
                  :ass2mouth:


Edited by trentallica (08/08/06 04:47 PM)


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