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Invisiblelipa

Registered: 07/24/07
Posts: 2,684
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: wbastz]
    #19216548 - 12/02/13 10:11 AM (10 years, 2 months ago)

Quote:

wbastz said:
Hi Terry,

I recently recovered a culture of Volvariella volvacea that was stored at room temperature in a screw cap test tube since 23/12/2001 (almost 12 years).

The mycelium was growing nicely on agar , but unfortunately I lost it in the next day due a failure of the thermostat of the incubator, when the temperature reached 59 ° C.

The mycelium was kept in distilled water , without any piece of agar, just a big cluster of mycelium. The tubes I use has the following dimensions: 100 x 15mm diameter with 4 ml of distiled water in every tube.

The same material stored on agar and on the same date of inoculation died.


Cheers,


Wilson




Thanks for the info. I use water agar before storage.


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OfflineShaggy Shaman
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19298455 - 12/19/13 03:18 PM (10 years, 2 months ago)

Just coming into this.

So I can sterilize these guys filled with distilled water, I'm assuming with the lids loose, then after cool down I can either drop an agar plug into the water or place the water onto the colonized petri, "stir" it, suck the solution up and squirt into the tubes, and let sit in mid 70's room temp for years?

Does the water ever need to be oxygenated? By shaking or anything?

If not, this sounds like the storage method I've truly been waiting for...

SS


--------------------
"Disbelief in magic can force a poor soul into believing in government and business."
— Tom Robbins

“The true value of a human being can be found in the degree to which he has attained liberation from the self"
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OfflineTerry M
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Shaggy Shaman]
    #19298724 - 12/19/13 04:28 PM (10 years, 2 months ago)

Hello Shaggy,

I use small screw-top vials. I sterilize these filled with distilled water and with the caps tight, just as you would for slants. Then, under sterile conditions, I open the cap and add the culture in some form: agar plugs, loose mycellium, etc. Then I screw the cap on tightly.

You don't allow them to breath like you would slants because in the sterile distilled water there are no nutrients, so nothing to metabolize and require air for. The mycellium goes into a sort of suspended animation. If you're lucky, it will wake up after some years.

I've tried both colonized agar plugs and loose mycelium. They both work, and I don't know if one is better than the other. Either way, the cultures are a pain to fish out of the SDW. I would maybe use this method as a last resort after slants. It can't be relied on 100%, but if you mess up your slants somehow (refrigerate tropical cultures, leave the cap screwed tight and forget to loosen it and seal with parafilm, allow a contaminant to sneak in), it could save you. And storage isn't a problem because you just put them somewhere at room temperature.


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OfflineShaggy Shaman
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Terry M]
    #19300079 - 12/19/13 09:11 PM (10 years, 2 months ago)

Thanks, Terry.

Since they aren't 100% reliable, doing multiple vials with the same genetics seems smart to me. As for fishing out the material later, why not plop the whole contents into an LC?

Picking your brain, here.


--------------------
"Disbelief in magic can force a poor soul into believing in government and business."
— Tom Robbins

“The true value of a human being can be found in the degree to which he has attained liberation from the self"
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OfflineJMcDoogle
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Shaggy Shaman]
    #19300446 - 12/19/13 10:50 PM (10 years, 2 months ago)

Sure am glad to see I've actually added something to these
forums that may after-all benefit someone!

:fuckinawesome:


--------------------
The ego is nothing other than the focus of conscious attention.


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Invisiblelipa

Registered: 07/24/07
Posts: 2,684
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Shaggy Shaman]
    #19302277 - 12/20/13 10:47 AM (10 years, 2 months ago)

Quote:

Shaggy Shaman said:
As for fishing out the material later, why not plop the whole contents into an LC?

Picking your brain, here.




One awesome way to create first generation spawn without having do a bunch of prep work every time is to do a session where you expand a lot of mycelium of a certain strain on weak nutrient agar and then throw a bunch of wedges into say 100 strait small plastic PPtubes and store. When you need to start expanding it just grab a tube and pour into sterilized grain and expand from there. They usually stay good for at least 6 month to a year before they poop out. Whats also cool is that they can be stored at room temperature.

Lipa


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Offlineforrest
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19302347 - 12/20/13 11:07 AM (10 years, 2 months ago)

you mean only a wedge in a tube, without additional water?
and it won't die by lack of oxygen?


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Invisiblelipa

Registered: 07/24/07
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: forrest]
    #19305690 - 12/20/13 11:31 PM (10 years, 2 months ago)

No.. there is water in the tube with the wedge.


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OfflineJMcDoogle
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19307785 - 12/21/13 12:16 PM (10 years, 2 months ago)

:smbfacepalm:


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The ego is nothing other than the focus of conscious attention.


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Offlineforrest
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: JMcDoogle]
    #19307967 - 12/21/13 01:02 PM (10 years, 2 months ago)

why the facepalm?

i know the thread is about storage in water, but for multiple years.

when someone gives a description of what to do but doesn't mentions to add water, and says the culture will stay good for 6 months to a year, it's not strange to think it is about something different than what the topic is about, and ask for clarification. but i guess the main difference in longivity is the low nutrient agar in stead of no nutrientss at all.

but i guess it's just stupid to ask questions, and i should go around giving people facepalms to feel better about myself.


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Invisiblefastfred
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: forrest]
    #19310151 - 12/22/13 12:55 AM (10 years, 2 months ago)

There's plenty of storage techniques out there.

Overlaying with mineral oil or adding a cryoprotectant and freezing are popular methods.

Mineral oil limits the oxygen available, thus reducing the metabolic rate.

Cryoprotectants allow you to freeze the culture, allowing you to store it essentially forever.  Milk and glycerin are popular cryoprotectants, so there's nothing fancy involved.


-FF


--------------------
It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy

The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed

"If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP)

I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid


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OfflineShaggy Shaman
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: fastfred]
    #19320247 - 12/24/13 09:23 AM (10 years, 2 months ago)

Those Argos cryovials are sold out, FYI. Anyone know where else I could get some without having to spend $400?

SS


--------------------
"Disbelief in magic can force a poor soul into believing in government and business."
— Tom Robbins

“The true value of a human being can be found in the degree to which he has attained liberation from the self"
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OfflineJMcDoogle
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Shaggy Shaman]
    #19320277 - 12/24/13 09:33 AM (10 years, 2 months ago)

You could try using a variation in ML -

Here are the 2 ML Tubes.

http://www.neobits.com/argos_technologies_cry12c_internally_threaded_cryovial_p5018690.html


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OfflineShaggy Shaman
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: JMcDoogle]
    #19322587 - 12/24/13 06:59 PM (10 years, 2 months ago)

2 mL seems kinda small, but I'll try them out if they have them.

Thanks, guys!

SS


--------------------
"Disbelief in magic can force a poor soul into believing in government and business."
— Tom Robbins

“The true value of a human being can be found in the degree to which he has attained liberation from the self"
-- Albert Einstein


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InvisibleAleon
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19328183 - 12/26/13 10:02 AM (10 years, 1 month ago)

Quote:

lipa said:
Quote:

Shaggy Shaman said:
As for fishing out the material later, why not plop the whole contents into an LC?

Picking your brain, here.




One awesome way to create first generation spawn without having do a bunch of prep work every time is to do a session where you expand a lot of mycelium of a certain strain on weak nutrient agar and then throw a bunch of wedges into say 100 strait small plastic PPtubes and store. When you need to start expanding it just grab a tube and pour into sterilized grain and expand from there. They usually stay good for at least 6 month to a year before they poop out. Whats also cool is that they can be stored at room temperature.

Lipa




Besides the fact that they can be stored at room temp, why is this adventageous over using petri dishes?  Just make a couple, throw them in the culture fridge, whenever u need a wedge take it out, transfer, then put back into fridge.  My petris last 1-2 years in fridge.


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OfflineJMcDoogle
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Aleon]
    #19328274 - 12/26/13 10:33 AM (10 years, 1 month ago)

Small footprint? No need for refrigeration - longer lifespan
Works with cultures that are cold sensitive, and heat sensitive, and hundreds of them can easily be stored in the same space as a normal fridge drawer.


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OfflineIcyus
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: JMcDoogle]
    #19328278 - 12/26/13 10:35 AM (10 years, 1 month ago)

Dont need that with your red spotty one..


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And thus begins the  reverse-fusing of our one-dimentional understanding, and adds ever-expanding perspectives, in depth and number; splitting our perception, and in so doing, seemingly irrationally, creates yet more one-ness, with all that ever was, is and will ever be, streching across the infinite, inunderstood concept of everything, percievable and not.


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Invisiblelipa

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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Aleon]
    #19330420 - 12/26/13 08:16 PM (10 years, 1 month ago)

less work. I can make up 100 tubes pretty quick. All year I can just make a G1 jar without having to do any petri work. Yes less clutter in the lab and more room in the fridge for masters. I only do it with common varieties that I know I will be using a lot.

Lipa


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InvisibleAleon
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19332226 - 12/27/13 09:13 AM (10 years, 1 month ago)

Im not really sure how its less work; but if it works for you, do it.  You make less transfers and the same amount of agar using the petri method as this water method. Instead of having to go petri->vial->spawn; I just go petri->spawn; eliminating a whole step (not to mention having to sterilize your water and those 100 vials in addition to your agar.) Adding extra steps adds extra vectors for contamination.  I do 1 agar transfer to make spawn; this method uses 2.  Instead of only having 1 opportunity to contaminate via transfers, this method has 2.  Also most growers have devoted culture fridges and a stack of petri dishes shouldn't be too much to fit into one.


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OfflineJMcDoogle
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Aleon]
    #19334876 - 12/27/13 11:02 PM (10 years, 1 month ago)

Now I dont agree with everything yo said but, this isnt an argument, its personal preference ya know. Nobody here has said its better than anything else, we're just
trying to be informative.


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