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OfflineTyperwritermonky
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Could One Make A Liquid Culture With Mycelium Rather Then Spores?
    #26714980 - 06/02/20 07:38 PM (3 years, 8 months ago)

Following the LC recipes to create one using spore syringes, could you not inject a solution of mycelium and sterelized water into the jar that the spores would otherwise be injected into?  Would this not let that mycelium prosper in the jar the same way and later be taken out with other syringes to then inject into bags?

Sorry if this is a very dumb question!


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OfflineMH5109
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Typerwritermonky] * 1
    #26714984 - 06/02/20 07:39 PM (3 years, 8 months ago)

Yes.


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OfflineTmethylM
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: MH5109]
    #26714992 - 06/02/20 07:44 PM (3 years, 8 months ago)

If you're very careful, maybe. The chances of transferring mycelium into an LC without contamination are small.


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Edited by Tmethyl (06/02/20 07:46 PM)


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Tmethyl] * 1
    #26715139 - 06/02/20 08:55 PM (3 years, 8 months ago)

Yes.

If your in an SAB or in front of a FH you have a better chance for clean LC using agar myclieum culture than you do spore.

Spores are dirty by the nature of fruiting. We fruit in open air so spores are going to be more prone to have an airborne contamination than a wedge of agar in a sealed dish.


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InvisibleStipe-n CapMDiscord
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Typerwritermonky] * 1
    #26715154 - 06/02/20 09:02 PM (3 years, 8 months ago)

like let's say....inoculate an LC with an LC ; )

What you're asking is pretty much how it's done other than the bit about sterile water. You can make LI to noc up an LC but you might as well just use a wedge because of what was mentioned above by Tmethyl, increased risk of contamination due to all of exposure in the steps to get there.

Any "recipe" that suggests to use spores to inoculate liquid culture should be ignored. Use clean mycelium from a plate or from a known clean LC (which I have done many times)


Edited by Stipe-n Cap (06/02/20 09:10 PM)


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OfflineIzzamagic
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Typerwritermonky]
    #26715165 - 06/02/20 09:05 PM (3 years, 8 months ago)

Quote:

Typerwritermonky said:
Following the LC recipes to create one using spore syringes, could you not inject a solution of mycelium and sterelized water into the jar that the spores would otherwise be injected into?  Would this not let that mycelium prosper in the jar the same way and later be taken out with other syringes to then inject into bags?

Sorry if this is a very dumb question!




Look up jose's biopsy lc method

Quote:

Tmethyl said:
If you're very careful, maybe. The chances of transferring mycelium into an LC without contamination are small.





Can you break down how this would be more likely to get contaminated as compared to a spore solution? My understanding is that unless prints are taken from invitro fruits, they are inevitably contaminated to some degree.


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InvisibleStipe-n CapMDiscord
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic]
    #26715170 - 06/02/20 09:07 PM (3 years, 8 months ago)

Exposure


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OfflineIzzamagic
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Stipe-n Cap] * 1
    #26715187 - 06/02/20 09:15 PM (3 years, 8 months ago)

Quote:

p9hu7 said:
Exposure




I'm struggling to see how sucking up spores and shooting them into an LC risks more exposure than poking a seemingly clean agar plate with a needle and doing the same. The process seems the same, just the source is different. I would assume a culture on agar after a few transfers would be a cleaner source


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OfflineTmethylM
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic] * 1
    #26715193 - 06/02/20 09:20 PM (3 years, 8 months ago)

Spores are sealed in a container, and injected into a sealed container.
1 vector of contamination, when the needle enters the LC through and injection port.

You have to open a plate of mycelium to open air, transfer the mycelium from the plate to the LC, through open air, the LC is also exposed to open air.
3 vectors of contamination.

Without a flowhood this would be nearly impossible.


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OfflineSockadin
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Tmethyl] * 1
    #26715214 - 06/02/20 09:32 PM (3 years, 8 months ago)

Yeah but to get said spores unless you are using a centerfugue the spores came from a print that fruited in open air so they have a high probability that they have a contaminate.

Verses myclieum from an agar culture.


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic]
    #26715219 - 06/02/20 09:33 PM (3 years, 8 months ago)

:whathesaid:

It should be obvious that spores are only known to be clean after the fact and they seldom are, where as cultures are known to be clean before inoculation.

Not only that but you specifically mentioned using sterile water with suspended mycelium. The extra steps from mycelium to sterile water to create liquid inoculant, then LI to LC are all risky and unnecessary exposure to possible contamination, as Tmethyl said earlier it's possible but....


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Offlinemarvins
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Tmethyl]
    #26715222 - 06/02/20 09:35 PM (3 years, 8 months ago)

Quote:

Tmethyl said:
Spores are sealed in a container, and injected into a sealed container.
1 vector of contamination, when the needle enters the LC through and injection port.

You have to open a plate of mycelium to open air, transfer the mycelium from the plate to the LC, through open air, the LC is also exposed to open air.
3 vectors of contamination.

Without a flowhood this would be nearly impossible.




I have done similar things (open LC transfer to syringe to open lid grain)in a SAB with sketchy sterile tek and have success so i think is obviusly possible.


Edited by marvins (06/02/20 09:37 PM)


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InvisibleStipe-n CapMDiscord
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: marvins]
    #26715231 - 06/02/20 09:38 PM (3 years, 8 months ago)

Nobody is saying it's impossible, what we are saying is that it's not the best practice. You can get away with it sometimes with low volume,  crank up your volume and your failure rate will be correlative. It's not really worth the risk, it will ultimately not pay off in the long run:shrug:

Using a SAB is not open air, just saying.


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OfflineIzzamagic
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Tmethyl]
    #26715247 - 06/02/20 09:44 PM (3 years, 8 months ago)

Quote:

Tmethyl said:
Spores are sealed in a container, and injected into a sealed container.
1 vector of contamination, when the needle enters the LC through and injection port.

You have to open a plate of mycelium to open air, transfer the mycelium from the plate to the LC, through open air, the LC is also exposed to open air.
3 vectors of contamination.

Without a flowhood this would be nearly impossible.




How did those spores get in that sealed container? Was the mushroom grown in an aseptic (invitro) environment and transferred to a sterilized container under a flowhood then sterilized water was added before being aspirated?

josex's biopsy method. agar poked with a needle/syringe containing sterilized water, injecting to broth either by cracking lid or through SHIp, all done in a SAB

https://www.shroomery.org/forums/showflat.php/Number/24740168#24740168


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OfflineTedTheHighlighter
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Tmethyl] * 1
    #26715257 - 06/02/20 09:47 PM (3 years, 8 months ago)

Quote:

Tmethyl said:
Spores are sealed in a container, and injected into a sealed container.
1 vector of contamination, when the needle enters the LC through and injection port.

You have to open a plate of mycelium to open air, transfer the mycelium from the plate to the LC, through open air, the LC is also exposed to open air.
3 vectors of contamination.

Without a flowhood this would be nearly impossible.




Wait, I’m very confused by this because I’ve seen other TCs like Bod say that using spores to inoculate LC is noob stuff and inoculating with agar is always preferred


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: TedTheHighlighter] * 1
    #26715264 - 06/02/20 09:49 PM (3 years, 8 months ago)

Yeah he's saying don't do it.

Actually, I don't know what the fuck is going on anymore lol.

OP just do biopsy syringe or transfer from a plate, spores>LC are dumb. Don't make LI to make LC, just make LC...


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Offlinemarvins
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Stipe-n Cap]
    #26715267 - 06/02/20 09:52 PM (3 years, 8 months ago)

Ah ok thats a good point i have only do 4 jars per session...
Quote:

p9hu7 said:
Using a SAB is not open air, just saying.




People works in open air? well as i was typing it i thought about working with big numbers in a SAB...:rolleyes:


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OfflineRapjack
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: marvins]
    #26715312 - 06/02/20 10:14 PM (3 years, 8 months ago)

It's doable, and preferred. Bodhi's posted pics of him using wedges for his LC's and the biopsy TEK seems good. That being said, a lot of TC's steer clear of LC altogether because it's impossible to guarantee cleanliness without first putting some drops onto agar which can contaminate it as well. It doesn't really seem to be a method often employed by commercial gourmet growers either, I think they find it simpler to just make a bunch of for sure clean agar plates and drop them onto grain.

Have you considered liquid inoculant (LI)? I've used it quite a few times successfully, it has a little bit of a learning curve to get the blend just right for fastest recovery but I find that it's worth it to reduce the number of times (and chances of contam) I have to do something to the culture and it cuts out the guesswork entirely  by using a clean plate.


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OfflineIzzamagic
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Rapjack]
    #26715318 - 06/02/20 10:17 PM (3 years, 8 months ago)

Quote:

Rapjack said:
It's doable, and preferred. Bodhi's posted pics of him using wedges for his LC's and the biopsy TEK seems good. That being said, a lot of TC's steer clear of LC altogether because it's impossible to guarantee cleanliness without first putting some drops onto agar which can contaminate it as well. It doesn't really seem to be a method often employed by commercial gourmet growers either, I think they find it simpler to just make a bunch of for sure clean agar plates and drop them onto grain.

Have you considered liquid inoculant (LI)? I've used it quite a few times successfully, it has a little bit of a learning curve to get the blend just right for fastest recovery but I find that it's worth it to reduce the number of times (and chances of contam) I have to do something to the culture and it cuts out the guesswork entirely  by using a clean plate.




touche! check out blenderless LI using the blenderbottle classic with that metal shaker peice in it. the classic is pressure cookable (with the lid loose). mix your agar softer to make breaking it up easier


Edited by Izzamagic (06/02/20 10:18 PM)


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Rapjack]
    #26715338 - 06/02/20 10:26 PM (3 years, 8 months ago)

Why is it that every noob mentions bod and "his"teks almost immediately.

Many TCs use LC and even if they didn't it's irrelevant. People do what they prefer personally like anyone else. There's definitely something to be said for the 5-7 day colonization with LC as apposed to 2-3 weeks with a2g.

If you'd like to make LC go here.
https://www.shroomery.org/forums/showflat.php/Number/26022486

Either make LC or make LI, no need to combine the two.
Spores to LC is el retardo...


Edited by Stipe-n Cap (06/02/20 10:40 PM)


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