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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, 7 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, 7 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, 7 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, 7 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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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Typerwritermonky] * 1
    #26715154 - 06/02/20 09:02 PM (3 years, 7 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, 7 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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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic]
    #26715170 - 06/02/20 09:07 PM (3 years, 7 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, 7 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, 7 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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¯\_(ツ)_/¯


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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, 7 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, 7 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, 7 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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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: marvins]
    #26715231 - 06/02/20 09:38 PM (3 years, 7 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, 7 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, 7 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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The cat asked, “Where do you want to go?”

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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, 7 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, 7 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, 7 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, 7 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, 7 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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OfflineLogicaL ChaosM
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Typerwritermonky]
    #26715815 - 06/03/20 05:32 AM (3 years, 7 months ago)

I would recommend transfering mycelium to agar, cleaning it up and then let it grow then add pieces of colonized agar to a liquid nutrient solution to make a Liquid Culture.


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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
    #26715880 - 06/03/20 06:05 AM (3 years, 7 months ago)

Quote:

p9hu7 said:
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...




How many times have you changed your mind just in this thread? Calm down


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Offlinepoisoned
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic] * 1
    #26715887 - 06/03/20 06:09 AM (3 years, 7 months ago)

Inoculating with spores is a big no-no.


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: poisoned] * 1
    #26715933 - 06/03/20 06:43 AM (3 years, 7 months ago)

Cuz Bodwin collected the good teks all in one spot, probably added some of his own twists, and made a flow chart. Everyone loves a flow chart bro. Now all the info you could ever need to grow tons of fun is right there and it’s so easy to just hit someone with a link to that and tell them to read read so it’s become like cube growing 101 mandatory no-ledge. Then the salty or standoffish attitude creates the barrier of mystique and draws more people in, seeking app. I watch all this stuff with fascination. People watching in the digital age, tinted by certain shared proclivities. A grand adventure we’re all on with this crazy spinning and swirling rock whose ephemeral shield protects us from the chaos. Proselytize


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Roger Clemency]
    #26715987 - 06/03/20 07:22 AM (3 years, 7 months ago)

I enjoy your observations. Are you enlightened? Cuz you sound enlightened. Can I follow you around? Maybe an address for donations?


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: LadysKnight]
    #26716002 - 06/03/20 07:30 AM (3 years, 7 months ago)

Quote:

LadysKnight said:
I enjoy your observations. Are you enlightened? Cuz you sound enlightened. Can I follow you around? Maybe an address for donations?




He’s in jail.
Send bail money.


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: ModularMind] * 1
    #26716013 - 06/03/20 07:34 AM (3 years, 7 months ago)

Quote:

ModularMind said:
Quote:

LadysKnight said:
I enjoy your observations. Are you enlightened? Cuz you sound enlightened. Can I follow you around? Maybe an address for donations?




He’s in jail.
Send bail money.




Directions unclear.
Sent hookers and blow.


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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: Izzamagic]
    #26716030 - 06/03/20 07:41 AM (3 years, 7 months ago)

Quote:

Izzamagic said:
Quote:

p9hu7 said:
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...




How many times have you changed your mind just in this thread? Calm down




Not once.

Quote:

Could One Make A Liquid Culture With Mycelium Rather Then Spores? 




Mycelium is used to make LC not spores

Quote:

could you not inject a solution of mycelium and sterelized water




Yes if it's a biopsy syringe to LC, or LC to LC, no if its LI because just make LC.

Pretty easy stuff.

Quote:

calm down








Edited by Stipe-n Cap (06/03/20 08:27 AM)


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OfflineDharmaForKarma
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: LadysKnight]
    #26716031 - 06/03/20 07:42 AM (3 years, 7 months ago)

Quote:

LadysKnight said:
Directions unclear.
Sent hookers and blow.




Now that is a magic 8 ball.


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OfflineTyperwritermonky
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Re: Could One Make A Liquid Culture With Mycelium Rather Then Spores? [Re: DharmaForKarma]
    #26716870 - 06/03/20 01:44 PM (3 years, 7 months ago)

Thanks yall for all the answers!  The liquid culture TEKs I found ALL had them being created by injecting a spore syringe into the sterilized sugar broth jar where they would prosper, so I was just thinking maybe it could be done with mycelium instead.

As well, this would be done with cyanescens (wavy caps) which are super hardy and tend to beat out other competition well.

Basically I have a jar of cyanescens mycelium with sterilized alder bark thats finally taken its foothold and is really starting to spread.  But the jar is too big and I am thinking of what I want to do with it to get things done in a timely manner.

So my idea is to get the mycelium onto agar and then use that to make a LC to grow directly from and inoculate with, and then for the rest of it put it into a smaller jar with better food and compost to make a fully colonized jar to then make outdoor beds with.

thanks yall for putting up with my ignorance!


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