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Sandala
Noob Shroomer

Registered: 02/20/20
Posts: 197
Loc: UK
Last seen: 1 year, 9 months
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help getting a clean culture?
#27076574 - 12/07/20 05:39 AM (3 years, 1 month ago) |
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I've been stuggling for best part of a year to get a clean culture, it looks clean after 4 or 5 transfers but then grows really slowly on oat grain, then doesn't survive first shake-up, just stops growing.
I've tried no pour plastic tubs, but these are a bit opaic and difficult to take pics of to show here, also no pour small jars but again difficult to show pics as the glass prevents getting a decent picture in focus.
I've purchased 200 petris to try to get this right, I'm giving up on the PDA, also giving up on oats will go for popcorn.
This is a pic showing the two distinctive types of growth on oats (presume the contam in the lower half of the pic has killed and outgrown the mycelium in the upper part of the pic) this is typical of my experience, this is 51 days since inoculation from a supplier bought LC syringe, no other contam showed up on agar from the syringe, in LC or direct to grain, only very weak tomatose growth. There is no contam/growth apart from where the LC went into the jars so it looks like the syringe brought this in with it rather than contam from my practices. Although it looks very similar to my previous multiple attempts from spore (2 different suppliers)

I've been using pre mixed powdered PDA, but I have yet to see any rhizomorphic growth, only very light tomatose growth, I would like an agar recipie that is a) more likely to show rhizomorphic growth, b) easier to separate or show distinctive difference between whatever this contam is and cube mycelium.
same efforts from agar

grain water LC, same culture

Any help or ideas what this contam might be, and a suggestion for agar recipe to combat the problem from spore would be much appreciated.
-------------------- My Noob Journal
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Camera93
We got dicks like Jesus



Registered: 08/15/18
Posts: 3,220
Last seen: 10 days, 12 hours
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Re: help getting a clean culture? [Re: Sandala]
#27076579 - 12/07/20 05:48 AM (3 years, 1 month ago) |
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however you gotta do it, get some pics of the plate
once it hits grain or LC it could be 100% cultivator error, but if the plates looks funky then you have a better shot at someone being able to help diagnose and give direction on to how to start cleaning the culture up
how are you prepping your oats? There is a lot of anecdotal experience with oats giving people mixed results. (any grain should work fine as long as you know how to properly hydrate and dry them)
What are you PC times? 2 hours 15 psi?
I have seen more spite towards popcorn then even oats around here btw lol so I'm not sure that change up is the best move have you given bird seed a shot? Fairly cheap depending on where you're located, and stupid simple to prep
-------------------- All I need are some tasty waves, a cool buzz, and I’m fine. Whatever you decide won’t really impact our survival Close your eyes, and do the best that you can
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Sunny Skies
Cluster Head


Registered: 05/03/17
Posts: 421
Loc: my house
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Re: help getting a clean culture? [Re: Sandala]
#27076584 - 12/07/20 05:52 AM (3 years, 1 month ago) |
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There is nothing wrong with PDA,its all I use and simple to make..ditch the LC and go from spore print.you may have better luck.
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Sandala
Noob Shroomer

Registered: 02/20/20
Posts: 197
Loc: UK
Last seen: 1 year, 9 months
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Re: help getting a clean culture? [Re: Sunny Skies]
#27076598 - 12/07/20 06:11 AM (3 years, 1 month ago) |
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I used Bods easy AF grain prep to the letter, 2hrs PC @15psi, I've done 6 or 7 batches now although, the first couple looked a bit too wet I made sure they were dry (no wet patch on paper towel) subsiquently.
to get a decent pic I have to take the lids off (not that it matter now I can't use these) this is typical of all my efforts...
these are old btw ~60+ days (holes in second pic from where transfers were taken)

The LC that went into the first pic (on the first post) was bought from a supplier, I was meticulous cleaning everything as i put it into the SAB, then cleaned again in the SAB, then sprayed iso up into the sSAB and let it rest for a couple of mins, then sprayed again up into the sab before doing anything, I was showered, gloved, masked and liberally wet with iso.
I have a spore print left, but it showed contam on the first couple of transfers, guess I'll just have to keep on trying when the new petri dishes arrive <shrug>
I feel like I'm pissing in the wind TBH
-------------------- My Noob Journal
Edited by Sandala (12/07/20 06:13 AM)
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Sunny Skies
Cluster Head


Registered: 05/03/17
Posts: 421
Loc: my house
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Re: help getting a clean culture? [Re: Sandala]
#27076678 - 12/07/20 07:40 AM (3 years, 1 month ago) |
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keep on trying with the print..those plates look like mould to me..how long did it take to germinate?? Have seen spores take 6 days or 3 weeks. Stay positive!
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The Mycologist
Explorer

Registered: 05/06/16
Posts: 3,024
Last seen: 29 days, 9 hours
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Re: help getting a clean culture? [Re: Sandala]
#27076724 - 12/07/20 08:17 AM (3 years, 1 month ago) |
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I wouldnt mess with LC until you are comfortable with pure agar work/inoculation.
I see BOD telling people this too.
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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Harry Manbach
Lock the door,Kill the light


Registered: 09/04/20
Posts: 339
Loc: 6
Last seen: 2 months, 22 days
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I'd certainly be taking transfers earlier too! You want to take samples from the leading edges to attempt to separate healthy mycelium from contaminants. It makes no sense to let your dishes over grow and sample from the middle where the contaminants are established!
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Sandala
Noob Shroomer

Registered: 02/20/20
Posts: 197
Loc: UK
Last seen: 1 year, 9 months
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This LC was purchased, I thought it would be an easy way to negate the issues, and start from a clean sample. I tried to make some myself earlier using the syringe biopsy method, it looked good but results turned out similar to the recent purchased LC.
All transfers were taken much earlier, from the leading edge, these pics are whats left months later.
10 days to germinate from spore, it's all in my journal, lots more pics, minus the latest purchased LC atempts.
I'll bin everything and try again when the petri dishes arrive, I'll do a lot more dishes and see how we go <shrug>
-------------------- My Noob Journal
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The Mycologist
Explorer

Registered: 05/06/16
Posts: 3,024
Last seen: 29 days, 9 hours
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Re: help getting a clean culture? [Re: Sandala]
#27077023 - 12/07/20 11:53 AM (3 years, 1 month ago) |
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I personally use no pour. Takes one vector out of the equation.
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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Sandala
Noob Shroomer

Registered: 02/20/20
Posts: 197
Loc: UK
Last seen: 1 year, 9 months
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Most of this was with no pour, trouble is I can't get a decent pic with the jars or the plastic tubs, and petri dish has a bigger surface area so it seems to give a better chance spotting contam <shrug>
-------------------- My Noob Journal
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Spirit-Crusher
DOOM HIPPIE


Registered: 04/24/19
Posts: 358
Loc: PNW
Last seen: 15 days, 15 hours
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Re: help getting a clean culture? [Re: Sandala]
#27078298 - 12/08/20 07:21 AM (3 years, 1 month ago) |
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Try switching grains. I hate oats.
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