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Invisiblelipa

Registered: 07/24/07
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Aleon]
    #19346508 - 12/30/13 02:14 PM (10 years, 1 month ago)

Quote:

Aleon said:
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.





I don't think you understand. The vials are made up the first time and only that time.  After that its vial -> grain every time for 6 months to a year.... I never get contamination doing it like this. A petri plate is more likely to contaminate in a fridge due to the fact that when you open it it is more exposed than a tiny vial. I prefer not to have to do any pre-culture work every time I have to make up a new generation.  Saves me time. I don't see how this is doing "more work".


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Offlinedrake89
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Registered: 06/26/11
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19346903 - 12/30/13 03:49 PM (10 years, 1 month ago)

You must make a lot of grain spawn (:

I finally figured out I only really need to make grain masters once per season, but I make a whole autoclave full of grain bags per species.


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Edited by drake89 (12/30/13 03:50 PM)


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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: drake89]
    #19347902 - 12/30/13 06:50 PM (10 years, 1 month ago)

Quote:

drake89 said:
You must make a lot of grain spawn (:

I finally figured out I only really need to make grain masters once per season, but I make a whole autoclave full of grain bags per species.





Yes, I sell spawn to local farms.  I g1-g2-bulk spawn or sub.  Always going to G2 at least to keep an eye on contamination. I am not trying to push the vial thing. It just works for me and I am just saying it works period.  I do the stay at home dad thing/spawn maker/mushroom grower. Every little time I can get is precious.  It's nice to be able to sterilize and just walk in there and plop a wedge in and done. No scalpel or any other tools needed.


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InvisibleAleon
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19350036 - 12/31/13 07:10 AM (10 years, 1 month ago)

Quote:

lipa said:
Quote:

drake89 said:
You must make a lot of grain spawn (:

I finally figured out I only really need to make grain masters once per season, but I make a whole autoclave full of grain bags per species.





Yes, I sell spawn to local farms.  I g1-g2-bulk spawn or sub.  Always going to G2 at least to keep an eye on contamination. I am not trying to push the vial thing. It just works for me and I am just saying it works period.  I do the stay at home dad thing/spawn maker/mushroom grower. Every little time I can get is precious.  It's nice to be able to sterilize and just walk in there and plop a wedge in and done. No scalpel or any other tools needed.





After you said THIS, now I see the convenience. That's nice not to have to bust out scalpel everytime u want to make masters; although I am on par with drake (I only make masters a few times a year anyways.) Also I wasn't trying to down-talk your method; I just didn't understand it fully.


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OfflineSnowArcher
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: Aleon]
    #19351165 - 12/31/13 01:24 PM (10 years, 1 month ago)

Thank you for this post.  I am recovering from a 3 year hiatus from growing (had to move to a very urban spot with no room) and am in the process of trying to revive my old library that is on slants and have had no attention.  Surprisingly, there are several that are already showing some signs of life.  It looks like 50%+ are dead though.  I will be using this method (and others) into the future.


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Re: Preserving cultures using sterile Distilled water in cryovials [Re: SnowArcher]
    #19377112 - 01/06/14 12:23 PM (10 years, 1 month ago)

would there be something against adding glycerine to agar/slant for storage in the fridge, to keep it from accidental freezing?


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Offlinematsc
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: forrest]
    #19382566 - 01/07/14 01:49 PM (10 years, 1 month ago)

I'm always wary of adding glycerol to things unless its going into the deep freeze (-80C). Some fungi can eat it and keep up some slow metabolic activity we want to avoid, and as it ages it tends to form all sorts of strange little peroxides that can damage the cells. In most cases, it would be probably be safe, but why risk it.

Its also a ROYAL PAIN IN THE ASS to pipette so I prefer to avoid it whenever possible (I may be a bit bitter here after spending a week making several hundred cryo stocks of our yeast and bacterial strains in lab. I was truly a defeated man by the end of that)


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Offlineforrest
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: matsc]
    #19533592 - 02/07/14 12:32 AM (10 years, 17 days ago)

thanks for that awnser matsc!

lipa, would 1,5 ml centifugetubes with myc scraped from agar, then with distilled water added so that (almost) no air is left when closing the lid, be the way to go about for the long term storage?


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Invisiblelipa

Registered: 07/24/07
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: forrest]
    #19534590 - 02/07/14 09:42 AM (10 years, 17 days ago)

Personally I use 5 mil sample tubes. Strait sides. The tubes are filled to 3/4 way full of distilled water and sterilized.  I run the fungus on water agar and then drop in about 4-5 small 3-4mm squares of the samples in each tube.  Then I set them right side up in a plastic container and set on the shelve in the lab. Just this week I pulled a few out that had 2008 written on them and they have already regrown out on to the agar. They were Lentinus squarrosulus and Lentinus giganteus.

You don't cool the tubes. They sit at room temperature undisturbed. No need for anything like glycerin etc.

Lipa


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Offlinedrake89
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: lipa]
    #19535068 - 02/07/14 12:13 PM (10 years, 17 days ago)

I will be giving this a go since it seems way cheaper and more fool-proof than glass slants :thumbup:


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Offlineforrest
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Re: Preserving cultures using sterile Distilled water in cryovials [Re: drake89]
    #19538009 - 02/08/14 03:05 AM (10 years, 16 days ago)

Thanks!


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Invisiblelipa

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Re: Preserving cultures using sterile Distilled water in cryovials [Re: drake89]
    #19538979 - 02/08/14 10:11 AM (10 years, 16 days ago)

Quote:

drake89 said:
I will be giving this a go since it seems way cheaper and more fool-proof than glass slants :thumbup:




I would make slants and use DWS for backup. I only pull them when the slants have some issues or they get neglected in some way. Its not full-proof and there is no way to monitor them except for using them. It is a valuable tool though. Especially for cultures that have sexual and asexual lifecycles (i.e. PL. cystidiosus.)

Lipa


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