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OfflinePsyence
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LC testing
    #26439866 - 01/18/20 08:58 AM (4 years, 29 days ago)

I don’t yet have the equipment to do agar work, but I have three jars of LC I made from a spore syringe from a good source.

Testing one on a batch of PFTek jars and so far 7 of 9 have shown signs of contamination. I have PC-Ed those ones again.

Question - if the LC is contaminated, would you expect every jar to be contaminated as a result? Is there a chance some would not?

As I said, 2 still look OK. Would you predict that they will also succumb to contamination?

Thanks


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InvisiblemushboyMDiscord
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Re: LC testing [Re: Psyence]
    #26439874 - 01/18/20 09:04 AM (4 years, 29 days ago)

yes and probably not.

contaminated lc will contaminate the universe. agar is a prerequisite for lc. doing lc without agar is like going into combat without ammo.


Edited by mushboy (01/18/20 09:06 AM)


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OfflinePsyence
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Re: LC testing [Re: mushboy]
    #26439892 - 01/18/20 09:17 AM (4 years, 29 days ago)

Awww :frown:

OK, thanks Mushboy.

Agar it is then!


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OfflinePsyence
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Re: LC testing [Re: Psyence]
    #26439901 - 01/18/20 09:20 AM (4 years, 29 days ago)

I had a bunch of grain prepped (soaked and PCed) Is there any way of saving that until I’m ready with agar (like for a week or two), or will I just need to toss it and start again?


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Re: LC testing [Re: Psyence]
    #26439919 - 01/18/20 09:30 AM (4 years, 29 days ago)

common best practice is to always use fresh grain jars but ive let jars sit for a long ass time and use them just fine.


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Re: LC testing [Re: Psyence]
    #26439921 - 01/18/20 09:31 AM (4 years, 29 days ago)

I've used pc'ed grain months old and was fine. If you do it right, you got time. Some people I know say to use it right out of the PC or certain doom. B.S. in my experience.


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OfflinePsyence
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Re: LC testing [Re: Smartattack]
    #26439923 - 01/18/20 09:33 AM (4 years, 29 days ago)

Good to hear!

I guess I could PC them again when I’m ready, or would that dry them out / cook them too much?


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Re: LC testing [Re: Psyence]
    #26439927 - 01/18/20 09:35 AM (4 years, 29 days ago)

why bother? pc new jars instead. now youll have twice as many sterilized jars!:rocket:


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Re: LC testing [Re: mushboy]
    #26470674 - 02/05/20 04:48 PM (4 years, 11 days ago)

Update: so I made myself a glovebox and agar gear, plus a borrowed microscope :smile:

I am currently trying 3 methods: agar culture from liquid culture, cloning from a mushroom and growing direct from spores. I have 4 questions:

All 3 of my plates from my LC are clean. Does that mean I can assume the LC itself is uncontaminated?

I am seeing much more rhizomorphic growth on one of the clone plates ( the other two look dead). Is this a better bet for grain spawn than the tomentose growth I am seeing on my LC plates or does it make no difference?

The spore plate is taking a long time to germinate. Is this normal? I just put the spores directly onto the agar.

If I grow clean on agar then transfer to a liquid medium, that means I will have a better chance of getting clean LC, right? I’m going to do more PFtek jars in addition to a grain spawn monotub, só for that I will need a liquid inoculant.

Thanks!


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OfflineA.k.aM
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Re: LC testing [Re: Psyence]
    #26470681 - 02/05/20 04:52 PM (4 years, 11 days ago)

Is the lc made from spores?

I’m pretty new to agar but I started with a bunch of clones and had lots of rhizo, I recently did a bunch of spore plates and they’re all much more thick and tomentose until a few transfers deep. Overall it doesn’t matter what kind of growth it is, you can manipulate it a lot with the agar recipes and plate thickness.

If the plates came up clean lc is clean.

When you have a clean agar plate cut a little wedge and put it in the lc.

Spores can take weeks to germinate. I started with spores Jan first and done prob 20 germ plates and they either had growth by a week or never went off, but I know people have had them take a month to germinate.


Edited by A.k.a (02/05/20 04:55 PM)


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Re: LC testing [Re: A.k.a]
    #26470726 - 02/05/20 05:11 PM (4 years, 11 days ago)

Thank you!

Yes, the LC I used was originally from spores, then transferred LC to LC . I made 3 jars, one of which is definitely contaminated. Weirdly the one I used for the agar (clean) was made from the contaminated one, so whether the contamination happened after that initial transfer I don’t know!

Is it necessary to do agar to agar transfers? Can you really speed up growth that way? Is speeding up growth the reason to do it?


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Re: LC testing [Re: Psyence]
    #26470763 - 02/05/20 05:27 PM (4 years, 11 days ago)

Necessary for what?

I think the main reason for agar is to make sure you have clean cultures, but I’ve definitely had slow ones that have sped up noticeably after transferring the fastest growth a few times.

I initially planned on using agar to expand myc for jars but recently did a g2g transfer for the first time and that’s 100% the way to go. Now I’m using it to get a clean plate to put into a half pint of seeds which will then be poured into multiple quarts.


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Re: LC testing [Re: A.k.a]
    #26470781 - 02/05/20 05:33 PM (4 years, 11 days ago)

Well, a lot of people seem to be doing sectoring, multiple transfers etc. Are they just doing it for fun? I guess the real necessity is identifying contaminants, but is there much to be gained by agar to agar transfers?

I’m going for a few jars of rye grain now. Don’t have the space to do more than one monotub yet, so no need for me to expand that rapidly yet!


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Re: LC testing [Re: Psyence]
    #26471391 - 02/06/20 02:38 AM (4 years, 11 days ago)

So my spore dish has germinated! Very pleased as this was a different strain (Hawaiian) to the rest (Albino A+).

Here’s another question: when selecting a mushroom for a spore print, I realise the selection of a particular specimen will not be as important as when cloning, but does it matter at all beyond size (so number of spores)?


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