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Offlinefranko142
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Why is spore syringe inoculation better than spore print inoculation?
    #25436658 - 09/05/18 04:26 AM (2 years, 8 months ago)

Hi,

I am wondering: Let's say you have fresh grow on your hands and you want to start a new cake based of the new grow...

From what I understand your options are (from best to worst):
1. Take spore prints -> inoculate agar with spores -> isolate single strain on agar -> inoculate with agar
2. Take spore prints -> create spore syringe(s) -> inoculate with syringe
3. Take spore prints -> inoculate with spore print

I read in other threads on this forum, that option 2 is far superior than option 3. What I don't understand is - why? I understand that prints aren't 100% sterile and that syringes are even less sterile than prints. So why would making a syringe increase success rate?

I understand that with a syringe you don't need to open the sterilized substrate container, but instead you can use a port. But option 1 is even better and you can't use a port just like with option 3.

Can someone please explain to me?

Also I heard that using pins instead of prints on agar is a very successful with mycelium outgrowing any potential contamination. Could you inoculate new substarte (brf/rye) with pins? Would that be single strain inoculation?

Some context: let's assume that number of jars/cakes you can inoculate doesn't matter as the goal is to inoculate a _single_ cake. So inoculating a single cake with a whole print is just as good creating 10 spore syringes, as only one is used anyways

EDIT: I am talking about ms syringes here. See for example this thread, where people are claiming print -> ms syringe -> inoculation is more successful than print -> inoculation:
https://www.shroomery.org/forums/showflat.php/Number/7983533
EDIT2: Just as an example here is a contradictory claim that print inoculation is superior: https://www.shroomery.org/forums/showflat.php/Number/7480618


Edited by franko142 (09/05/18 06:06 AM)


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Offlinebishlap
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142]
    #25436669 - 09/05/18 04:45 AM (2 years, 8 months ago)

I think your confusing ms syringe with live culture syringe.

Multi spore syringes are a crap shoot and with live culture you can inject live mycelium thereby expediting the inoculation process.


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Offlinefranko142
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: bishlap]
    #25436692 - 09/05/18 05:18 AM (2 years, 8 months ago)

Let me check if we are on the same page...

ms syringe: A syringe created by sucking up a spore print (+ water) (as outlined in option 2)
live culture syringe: A syringe created from a liquid culture, which consist of isolated mycelium + a nutrious liquid (e.g. honey in water). Not based on a spore print (not outlined in any option), but on agar? Tissue?

Is this correct?

In that case what does it mean?
- You would say a ms syringe is worse than a print?
- A live culture syringe is better than a print?
- How would you go about creating a live culture without agar and have less contamination than with a print if at all possible?

EDIT: Also in my first post I was talking about MS syringes all the way. I added a reference to my first post.


Edited by franko142 (09/05/18 05:29 AM)


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Offlinebishlap
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142]
    #25436697 - 09/05/18 05:33 AM (2 years, 8 months ago)

Ms syringes are made from prints. And aren't terrible, your almost guaranteed to get fruits and everyone needs to start somewhere it's just not likely your going to get a full carpet of mushies

There's plenty of teks in the search.
Agar is easy I have used tiny 1/2 pint jars as agar dishes and agar is cheap.

Your on the right path just do more research, the search engine on this site can answer almost any question.


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Offlinefranko142
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: bishlap]
    #25436701 - 09/05/18 05:45 AM (2 years, 8 months ago)

I did a lot of research and read a lot of teks already. I know about the search engine and used it quite extensively already. I even read the relevant parts of The Mushroom Cultivator. My problem now is not a lack of research, but contradicting, unintuitive and unexplained information.

I would appreciate it if someone could answer any or all of my questions instead of telling me to ask different questions. I know about agar and it's benefits (as my initial post in this thread shows), but I am asking specifically about details of non-agar teks compared to each other.


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OfflineThe-Doc
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142]
    #25436715 - 09/05/18 06:06 AM (2 years, 8 months ago)

Quote:

franko142 said:
I did a lot of research and read a lot of teks already. I know about the search engine and used it quite extensively already. I even read the relevant parts of The Mushroom Cultivator. My problem now is not a lack of research, but contradicting, unintuitive and unexplained information.

I would appreciate it if someone could answer any or all of my questions instead of telling me to ask different questions. I know about agar and it's benefits (as my initial post in this thread shows), but I am asking specifically about details of non-agar teks compared to each other.



Where did you read about inoculating directly from a spore print?


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Offlinefranko142
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: The-Doc]
    #25436720 - 09/05/18 06:12 AM (2 years, 8 months ago)

https://www.shroomery.org/forums/showflat.php/Number/586065
https://www.shroomery.org/forums/showflat.php/Number/7480618

I simply can't understand WHY a syringe would be superior to a print, I can't find any explanation on why. And in regards to whether prints to substrate work at all... there is a lot of contradicting information out there. Some say it works better (makes sense to me) and some say it's impossible/unlikely.


Edited by franko142 (09/05/18 06:21 AM)


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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142] * 1
    #25436750 - 09/05/18 07:13 AM (2 years, 8 months ago)

Quote:

franko142 said:
I did a lot of research and read a lot of teks already. I know about the search engine and used it quite extensively already. I even read the relevant parts of The Mushroom Cultivator. My problem now is not a lack of research, but contradicting, unintuitive and unexplained information.

I would appreciate it if someone could answer any or all of my questions instead of telling me to ask different questions. I know about agar and it's benefits (as my initial post in this thread shows), but I am asking specifically about details of non-agar teks compared to each other.



Doesn't sound like it but assuming its true look up only newer threads. See my search engine tek(in my links). And forget that old book. 1000s of people come hear each year and find no lack of research, little contradiction, and that things are very intuitive and extremely explained. But it takes good research skills otherwise yes you'll absolutely find it confusing.

I personally think prints are way superior to syringes. The question doesn't matter tho, once you know how to use them you'll be answering your own question. They're better for different things

People can isolate monocultures on agar but almost no one goes thru the hassle. You probably can't isolate a single strain on agar without fancy spore dilution techniques. You would want to get the single strain off the bat rather than try to separate it out of a culture.
Either way untested monocultures or single strains are a crapshoot. More people would rather just clone a mushroom and know that they have highly likely to be good genetics right off the bat.


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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: bodhisatta]
    #25436755 - 09/05/18 07:25 AM (2 years, 8 months ago)

Have you ever tried inoculating jars with a spore print and and exacto knife? It's not fun and has too many steps.
That's why a syringe is superior because of was if use and sterilization.

I have heard of people putting live mushroom tissue in a liquid culture but honestly man if your just starting out don't worry about advanced techniques right now they will become kinda common sense in the future.
And don't be afraid of agar it's really cool even if just to play around with.


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OfflineSolipsis
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: bishlap]
    #25436817 - 09/05/18 08:35 AM (2 years, 8 months ago)

Regarding the OP:

What is wrong about blindly isolating on agar is you may pick a poor isolate which can't even fruit properly. What is good about germinating on agar first is better ensuring that you have no contams.
But you can inoculate with a piece of agar when the multispore culture is not yet resolved into an isolate (something which may require multiple transfers anyway, I believe this depends on the spore concentration: less spores means easier resolution?). This should better ensure that in the MS culture there are good parts.

Comparing this to using spores either directly or with a syringe: if the print is not clean, making a syringe may increase the chance of suspending a contam in the process by a lot. On the other hand: if the print is clean, making a syringe allows hydration of the spores which gives them a headstart with germination.


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Offlinefranko142
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: Solipsis]
    #25436963 - 09/05/18 10:11 AM (2 years, 8 months ago)

@bishlap: What do you mean with "too many steps"? I'd say it has less steps than preparing a syringe or what am I missing?

I didn't consider the crap shit with isolated mycelium... thanks for the info.

What about inoculation with pins? Has anyone an opinion on that?


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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142]
    #25437008 - 09/05/18 10:40 AM (2 years, 8 months ago)

Pins should be cloned to agar. You're inoculating agar with the pin. Going right to grain would be a risk.


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Offlinedelcat
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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: bodhisatta]
    #25437028 - 09/05/18 10:52 AM (2 years, 8 months ago)

Quote:

bodhisatta said:
Pins should be cloned to agar. You're inoculating agar with the pin. Going right to grain would be a risk.




Just to add my uneducated elaboration to Bodhi's comment - going right to grain would be a risk, because the outside of the pin is exposed to air, and thus contams.

If you clone your pin to agar, you can use standard agar transfer techniques to make sure you have completely clean agar before you innoc grains from it.  Its still the same genetic material as the pin, you've just taken the time to ensure all the contams are out of there.

There really isn't a way to cut corners on sterile procedure.


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Re: Why is spore syringe inoculation better than spore print inoculation? [Re: franko142]
    #27028786 - 11/08/20 05:03 PM (6 months, 2 days ago)

Thank you for this post! Finally I’m reading good questions and answers in Laymans terms I can understand! The general info is so damn overwhelming to me.


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