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Invisiblealaskappalachian
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Registered: 10/22/19
Posts: 1,688
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Re: Instant pot for sterilization [Re: Mushroomsee]
    #26511103 - 03/01/20 09:39 AM (3 years, 11 months ago)

Sorry bud- I missed that you wanted to use bags.  You can seal them afterwards, but I just twist and use metal twist-tie to seal (much easier IMO)  You want to let those bags cool a good while before using becasue the core of the grain mass can stay hot for a while.  You want to inoculate @ <100f, as a genereal rule.  Some people pre-seal bags frist.  I have pre-sealed filter patch bags before (and my own bags made with food-saver bags and micropore tape layered for a "filter patch"), but on the rare occasion I use a bag now, I'm inocuating with agar or colonized grain, so it's adding an extra step.  It would seem you are planning to inject a spore solution into your grain bag.  I highly, highly reccomend getting some agar and making some pre-pours to clean up that dirty sore solution before inoculating your grain bags, btw.  Hate to see you go through all this trouble just to fail.  Multispore solution to grain is risky and ripe for a high rate of failure.  Ayeplus's Bag Tek is up on the first page of threads and gives you the basics btw.


Edited by alaskappalachian (03/01/20 09:52 AM)


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InvisibleMushroomsee
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Re: Instant pot for sterilization [Re: alaskappalachian]
    #26511221 - 03/01/20 11:11 AM (3 years, 11 months ago)

Quote:

alaskappalachian said:
Sorry bud- I missed that you wanted to use bags.  You can seal them afterwards, but I just twist and use metal twist-tie to seal (much easier IMO)  You want to let those bags cool a good while before using becasue the core of the grain mass can stay hot for a while.  You want to inoculate @ <100f, as a genereal rule.  Some people pre-seal bags frist.  I have pre-sealed filter patch bags before (and my own bags made with food-saver bags and micropore tape layered for a "filter patch"), but on the rare occasion I use a bag now, I'm inocuating with agar or colonized grain, so it's adding an extra step.  It would seem you are planning to inject a spore solution into your grain bag.  I highly, highly reccomend getting some agar and making some pre-pours to clean up that dirty sore solution before inoculating your grain bags, btw.  Hate to see you go through all this trouble just to fail.  Multispore solution to grain is risky and ripe for a high rate of failure.  Ayeplus's Bag Tek is up on the first page of threads and gives you the basics btw.




Damn.. this is just getting more and more advanced hehe :grin: I saw that you could do g2g too, as you said. What do you think about that instead of agar? I already have a rye bag that is pre-sterilized as well as a spore syringe. Wondering if I should use the syringe for that bag, and then run the g2g from that one to all my own made spawn bags I will make?

Thanks again mate,

mush love


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Invisiblealaskappalachian
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Registered: 10/22/19
Posts: 1,688
Loc: The 49th Dimension
Re: Instant pot for sterilization [Re: Mushroomsee]
    #26511251 - 03/01/20 11:30 AM (3 years, 11 months ago)

You could use a few ccs of that syringe and try out that bag, but like I said: injecting multispore solution into a grain bag is risky.  You are sitting on that bag, so you'll still have way, way mrorre than enough left to get some agar going iff oyu want to use a squirt and see if you get lucky with that bag.  You never know.  But I wouldn't coun't on it being anything you'd want to g2g with....  You really need to use agar to get a clean culture.  Really not advanced at all.  Cheap and super-easy.
https://www.shroomery.org/forums/showflat.php/Number/19208976&nbsp; You can use little 4oz or wide half pint jars too.


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OfflineFrankRhizo
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Re: Instant pot for sterilization [Re: alaskappalachian]
    #26511418 - 03/01/20 01:35 PM (3 years, 11 months ago)

.


Edited by FrankRhizo (03/01/20 01:36 PM)


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Offlinepatapon333
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Re: Instant pot for sterilization [Re: FrankRhizo]
    #26511426 - 03/01/20 01:41 PM (3 years, 11 months ago)

You can reach a sterile environment by Tyndallization - you need it to boil 15 min, wait 24 hours to the endospores start germinating and boil again. You need no pressure. Of course it is much easier with a pressure cooker for 90 min. For this hobby there is no better way than this.


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InvisibleMushroomsee
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Registered: 04/29/18
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Re: Instant pot for sterilization [Re: alaskappalachian]
    #26511512 - 03/01/20 02:43 PM (3 years, 11 months ago)

Quote:

alaskappalachian said:
You could use a few ccs of that syringe and try out that bag, but like I said: injecting multispore solution into a grain bag is risky.  You are sitting on that bag, so you'll still have way, way mrorre than enough left to get some agar going iff oyu want to use a squirt and see if you get lucky with that bag.  You never know.  But I wouldn't coun't on it being anything you'd want to g2g with....  You really need to use agar to get a clean culture.  Really not advanced at all.  Cheap and super-easy.
https://www.shroomery.org/forums/showflat.php/Number/19208976&nbsp; You can use little 4oz or wide half pint jars too.




Alright, many thanks again. I'll do exactly as you say and test a little in the bag I have, and take the rest to agar. Found what I need to do to make agar plates so I will give it a shoot. But one question, does it matter that I use the multispore syringe for agar? Will my agar be bad due to multispore, or is the agar that way still much better despite the multispore I use?


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Offlineantmanmax
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Re: Instant pot for sterilization [Re: Mushroomsee]
    #26511565 - 03/01/20 03:10 PM (3 years, 11 months ago)

Quote:

Mushroomsee said:
Alright, many thanks again. I'll do exactly as you say and test a little in the bag I have, and take the rest to agar. Found what I need to do to make agar plates so I will give it a shoot. But one question, does it matter that I use the multispore syringe for agar? Will my agar be bad due to multispore, or is the agar that way still much better despite the multispore I use?




It won't be "bad" per se, but most people strive to isolate monocultures for reliability. After a few transfers you'll usually end up with a monoculture.


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InvisibleMushroomsee
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Re: Instant pot for sterilization [Re: antmanmax]
    #26511588 - 03/01/20 03:22 PM (3 years, 11 months ago)

Quote:

antmanmax said:
Quote:

Mushroomsee said:
Alright, many thanks again. I'll do exactly as you say and test a little in the bag I have, and take the rest to agar. Found what I need to do to make agar plates so I will give it a shoot. But one question, does it matter that I use the multispore syringe for agar? Will my agar be bad due to multispore, or is the agar that way still much better despite the multispore I use?




It won't be "bad" per se, but most people strive to isolate monocultures for reliability. After a few transfers you'll usually end up with a monoculture.




Are there other benefits to it instead of just using the syringe straight into the rye bag? Does it colonize faster with agar than with spore syringe?


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InvisiblemushboyMDiscord
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Re: Instant pot for sterilization [Re: antmanmax]
    #26511592 - 03/01/20 03:25 PM (3 years, 11 months ago)

The culture can be cleaner than the spores. Clean myc colonizes really fast compared to bacterial/moldy inoculate. Plus the myc is already germinated:wink:

Quote:

antmanmax said:
After a few transfers you'll usually end up with a monoculture.




A few? More like 50+


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InvisibleMushroomsee
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Re: Instant pot for sterilization [Re: mushboy]
    #26511610 - 03/01/20 03:36 PM (3 years, 11 months ago)

Quote:

mushboy said:
The culture can be cleaner than the spores. Clean myc colonizes really fast compared to bacterial/moldy inoculate. Plus the myc is already germinated;)

Quote:

antmanmax said:
After a few transfers you'll usually end up with a monoculture.




A few? More like 50+




okay but if you skip the isolation, then the advantage of agar is only that it is already germinated? How much colonization time do you save from that?

Another advantage as I understood is that a spore syringe can be made into a lot of agar so that you can basically just continue using agar all the time and never buy new spore syringes? Right?


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Invisiblealaskappalachian
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Registered: 10/22/19
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Re: Instant pot for sterilization [Re: Mushroomsee]
    #26511613 - 03/01/20 03:38 PM (3 years, 11 months ago)

Your first plate (the plate you introduce a drop of spore solution to initially- your germ(ination) plate) will have some travellers on it, and will need to be transfered again a couple times at least to clean up.  Seems like a lot of work but when you have a clean culture you don't have to wait and watch as your bags go bad.  You can then colonize bags or jars of grain and keep doing g2g transfers to your heart's content.  Just take a little more time to read up on the hows and whys of agar. 

Read this- it's a good walkthrough with everything you need to know to get started with agar.  Best of luck!

https://www.shroomery.org/forums/showflat.php/Number/21176491


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OfflineA.k.aM
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Re: Instant pot for sterilization [Re: Mushroomsee]
    #26511616 - 03/01/20 03:39 PM (3 years, 11 months ago)

I think you’re misunderstanding agar.

It’s just something to grow the myc on so you can be sure there’s no contamination before you put it into a jar. Once you have mycelium on agar you could grow as much as you want until you run out of plates.


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Offlineantmanmax
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Re: Instant pot for sterilization [Re: mushboy]
    #26511629 - 03/01/20 03:51 PM (3 years, 11 months ago)

Quote:

mushboy said:
The culture can be cleaner than the spores. Clean myc colonizes really fast compared to bacterial/moldy inoculate. Plus the myc is already germinated;)

Quote:

antmanmax said:
After a few transfers you'll usually end up with a monoculture.




A few? More like 50+




Does it really take this many to get a monoculture? I've had good results after around a dozen.


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InvisiblemushboyMDiscord
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Re: Instant pot for sterilization [Re: antmanmax]
    #26511662 - 03/01/20 04:10 PM (3 years, 11 months ago)

Isolate*

It takes 50ish(a shitload) of transfers to get an isolate.

Monocultures can happen sooner but not every monoculture is an isolate I think.. could be wrong on that tho:shrug:


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Offlinejbgtaa
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Re: Instant pot for sterilization [Re: mushboy]
    #26511742 - 03/01/20 05:22 PM (3 years, 11 months ago)

A monoculture just technically means one kind of mycelium on the agar. Isolates are 1 pure dikaryon which are hard to achieve even with 50+ transfers

To clarify it’s possible to achieve a full isolate. Only problem is we as hobbyists don’t have the type of knowledge and experience to know exactly how to culture towards an isolate. There’s a good amount of people on this site who could achieve it but even then isolate doesn’t always mean a good fruiting culture.


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Edited by jbgtaa (03/01/20 05:24 PM)


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OfflineBarrel
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Re: Instant pot for sterilization [Re: Mushroomsee]
    #26512205 - 03/02/20 01:58 AM (3 years, 11 months ago)

Quote:

Mushroomsee said:
Also, in this video, the guy uses gypsum and coffee in the water with his rye grains. Is it needed? Does it make a big difference?

At 2.34-3.00





It was all the rage about a decade ago when cultivators were freaking out about pH and nitrogen content. It's really not needed for cubes, but some gourmet varieties do a bit better with supplements. It fell out of favor because it over complicates the process for marginal gains atbest.


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InvisibleMushroomsee
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Re: Instant pot for sterilization [Re: alaskappalachian]
    #26512906 - 03/02/20 01:47 PM (3 years, 11 months ago)

Thanks for all the answers and information guys, learning alot from you. I will proceed with the process.

Mush love


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