|
eugenemyco
Stranger



Registered: 01/30/23
Posts: 80
Last seen: 4 months, 25 days
|
Logistics of long term storage - help
#28365825 - 06/19/23 04:00 PM (1 year, 6 months ago) |
|
|
Currently working on a bunch of new plates in search of well-performing cultures that I want to then put to long-term storage.
I am currently testing out clone plates in search of these cultures.
I have backups of all the plates that I'm testing on grain, but those backups don't necessarily exhibit the same growth.
Not sure what the best way to go about saving the culture if the one that goes to grain performs well.
When/how would you guys go about transferring a well-performing culture to slant?
Need help with the logistics.
|
Baba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,496
Loc: Try 'n' Error Boot Camp
|
Re: Logistics of long term storage - help [Re: eugenemyco] 2
#28365876 - 06/19/23 04:43 PM (1 year, 6 months ago) |
|
|
So what exactly is your concern? Are you afraid you are loosing some of the genetic profile of the culture when taking transfers?
Sounds like a solid plan you got there. Culture to grain, backup culture on hold, once good performance is confirmed transfer to slant or preserve with other methods.
You could go about it in different ways, could even keep some of the colonized spawn and dry it out for later use like yoshi trainer and fahtster do or put one of the grains to a plate to preserve the culture. I personally inoculate everything with LC which I also use for long term storage.
I wouldn't get too hung up about the backup cultures looking a bit different as long as they are clean.
|
eugenemyco
Stranger



Registered: 01/30/23
Posts: 80
Last seen: 4 months, 25 days
|
Re: Logistics of long term storage - help [Re: Baba Yaga]
#28366182 - 06/19/23 07:33 PM (1 year, 6 months ago) |
|
|
Thanks, that's super insightful actually.
You've suggested a bunch of viable approaches so I'm far less concerned now.
I was originally concerned that the backups wouldn't perform the same as the plates that've gone to grain. In part due to the fact that i'm fairly new to agar work, that my transferring wouldn't be consistent (hence the differing growth between plates from the same transfer session).
Additionally I didn't know if taking colonized grain to agar was an established or experimental practice. Knowing that a few TC's do this is reassuring. Also didn't know that LC could used in more than one session out of caution for further contaminations, as I recall a tek advising to use LC once it's made. Though I can't remember which. I suppose it all comes down to sterile technique.
I am always curious about when and how others put their cultures to storage, and for what scale of grow they operate.
|
bongoman
Stranger

Registered: 09/19/18
Posts: 579
Last seen: 3 days, 1 hour
|
Re: Logistics of long term storage - help [Re: Baba Yaga]
#28366320 - 06/19/23 09:15 PM (1 year, 6 months ago) |
|
|
Quote:
Baba Yaga said:I personally inoculate everything with LC which I also use for long term storage.
Can I ask what type of containers you are storing your LC in for longer-term storage?
|
Baba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,496
Loc: Try 'n' Error Boot Camp
|
Re: Logistics of long term storage - help [Re: bongoman] 1
#28366368 - 06/19/23 10:11 PM (1 year, 6 months ago) |
|
|
Quote:
eugenemyco said: [...] didn't know that LC could used in more than one session out of caution for further contaminations, as I recall a tek advising to use LC once it's made. Though I can't remember which. I suppose it all comes down to sterile technique.
Yeah, it pretty much simmers down to sterile technique. Some people use the same LC in more than one sessions. I don't though and prefer making a few smaller LC and keep 1 or 2 as backup, if I ever run low I expand an LC to new broth before using it for grain but only if I really like the culture. TBH I'm just having fun playing around and get rarely attached to a certain culture, got more stashed than I can make use of anyway so this puts things into a different perspective.
Quote:
bongoman said:
Quote:
Baba Yaga said:I personally inoculate everything with LC which I also use for long term storage.
Can I ask what type of containers you are storing your LC in for longer-term storage?
Using 100 and 250 ml media bottles for LC which I only fill 2/3. They do act more like liquid plates and 60 ml can inoculated 10 jars. Never understood why hobby growers would make 500ml of LC. I screw the lid shut and wrap the neck with glad wrap to keep things clean. Storing at room temp.
|
eugenemyco
Stranger



Registered: 01/30/23
Posts: 80
Last seen: 4 months, 25 days
|
Re: Logistics of long term storage - help [Re: Baba Yaga] 1
#28367565 - 06/20/23 05:18 PM (1 year, 6 months ago) |
|
|
Baba Yaga thanks for putting me on, those 100ml and 250ml media bottles are so sick, I don't know what it is but I just really like them. Think I will start my LC journey with those.
Do you allow your LC to colonize with the cap slightly ajar, and then seal tight and wrap?
|
Baba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,496
Loc: Try 'n' Error Boot Camp
|
Re: Logistics of long term storage - help [Re: eugenemyco]
#28367620 - 06/20/23 06:12 PM (1 year, 6 months ago) |
|
|
The LC broth I make is 0.1% LME. Usually I fit 60ml into the 100ml bottles and about 120ml into the 250ml bottles. Once they are inoculated I screw the lid down tight and wrap the neck. I do shake the ones I am inoculating with more often and leave the ones I plan to use for storage more or less alone so they grow out in only a few thicker colonies. There is more than enough air in there to let them grow out and at some stage they'll go dormant.
|
NFLProof
Sponge



Registered: 05/31/18
Posts: 922
Loc: In the ether
Last seen: 25 days, 15 hours
|
Re: Logistics of long term storage - help [Re: Baba Yaga]
#28367664 - 06/20/23 06:52 PM (1 year, 6 months ago) |
|
|
Would there be a benefit to store the backup LCs in the fridge as well to persevere further as long as we protect against temperature swings?
-------------------- Yeah, yeah.
|
Baba Yaga
Psychedelic Minion

Registered: 09/13/20
Posts: 4,496
Loc: Try 'n' Error Boot Camp
|
Re: Logistics of long term storage - help [Re: NFLProof]
#28368085 - 06/21/23 01:39 AM (1 year, 6 months ago) |
|
|
IDK TBH. I don't have much experience with this and two years is the longest I have gotten so far except from an oyster LC which I had sitting in the fridge for 7 years and which bounced back instantly.
Personally I'd like to avoid the fridge and this is why I'm not testing cold storage. My guess is that if you get the cultures to switch into hibernation/dormancy then they will keep for a long time at stable room temperature. Cultures kept in distilled water is working for a very long time at room temperature according to some papers (google: "castellani long term storage of fungi" or similar).
I hope the low nutrient content of 0.1% will just do the same or maybe pouring some of the LC into distilled will improve longevity. I'd be happy with 3-4 years and then refresh by pouring some into fresh broth. Doing this with LC just fits my workflow and doesn't cause extra work, all I have to do to keep a culture is to put one of the LCs aside.
One day I will do trials with cultures on filter paper to store them dry.
|
NFLProof
Sponge



Registered: 05/31/18
Posts: 922
Loc: In the ether
Last seen: 25 days, 15 hours
|
Re: Logistics of long term storage - help [Re: Baba Yaga]
#28368342 - 06/21/23 07:55 AM (1 year, 6 months ago) |
|
|
Two years at room temperature is plenty in terms of long term for myself as well. Thank you.
-------------------- Yeah, yeah.
|
|
|
You cannot start new topics / You cannot reply to topics HTML is disabled / BBCode is enabled
Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, hamloaf, cronicr, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a 492 topic views. 22 members, 96 guests and 79 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ] |
|