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OfflineNoobShroob
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Mushroom to sterile grain to spawn?
    #28152685 - 01/22/23 07:01 PM (1 year, 2 months ago)

If i put a fresh grown mushroom into a sterilized grain bag would it spawn?

The plan would be to careful cut open a sterilized bag, drop a mushroom inside and then seal the bag.

I could also make an lc and inject, it does have a injection port so let me know what yall would think to be better. I dont have a sterilized needle but MIGHT be able to throw it in a pc, if not maybe alcohol and a lighter lol, let me know.


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OfflineGreg
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob] * 1
    #28152692 - 01/22/23 07:06 PM (1 year, 2 months ago)

An invitro pin from an agar plate would probably work but it would be pointless when you could just a2g instead.
A fruit from a normal grow is gonna have some amount of nasties on it no matter how clean your environment is, just like with spore prints / swabs / syringes.

Not sure how the LC question is related to the fruit to grain question, you might need to provide more info.
Verifiably clean LC to grain is fine, fruit to LC to grain would be even worse than fruit to grain.

Edit:
You might be able to take tissue from inside a fruit like you're taking a clone and drop that in either grains or LC.
I would just take a clone to agar first in that case though.

Edited by Greg (01/22/23 07:14 PM)

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Invisiblebakedbeings
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Re: Mushroom to sterile grain to spawn? [Re: Greg] * 1
    #28152708 - 01/22/23 07:17 PM (1 year, 2 months ago)

faht does something like what youre describing

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OfflineNoobShroob
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Re: Mushroom to sterile grain to spawn? [Re: Greg]
    #28152757 - 01/22/23 07:47 PM (1 year, 2 months ago)

Quote:

Greg said:
Not sure how the LC question is related to the fruit to grain question, you might need to provide more info.
Verifiably clean LC to grain is fine, fruit to LC to grain would be even worse than fruit to grain.
Edit:
You might be able to take tissue from inside a fruit like you're taking a clone and drop that in either grains or LC.
I would just take a clone to agar first in that case though.




Its related because im trying to find the best method with options that are available.

Tou say it would be worse to make it an lc and then inject but isnt that the whole point on an lc. Hey if you say its worse than i hear you but why bother with an lc at all then?

Quote:

bakedbeings said:
faht does something like what youre describing





That looks a lot more indepth/complicated than what i had in mind lol


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OfflineGreg
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob] * 1
    #28152766 - 01/22/23 07:51 PM (1 year, 2 months ago)

LC is great! I use LC all the time.
However, LC can be easy to screw up with less than stellar technique.
It can also hide contaminants, especially bacteria, pretty well.

It's a liquid, the tiniest current can expose a contaminant organism to every milliliter of it and you may not know until you test it.
Even worse if the organism is motile (has the ability to swim) like many bacteria do.

This is a video of a bacteria infested liquid culture I made years ago that looked crystal clear to the naked eye, taken with my microscope:


I didn't mean to imply LC as a whole is a bad idea, just that taking potentially dirty whole mushrooms and inoculating LC broth with them could easily result in a bad time.

Edited by Greg (01/22/23 07:54 PM)

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OfflineB Traven
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Re: Mushroom to sterile grain to spawn? [Re: bakedbeings]
    #28152782 - 01/22/23 08:03 PM (1 year, 2 months ago)

If I run my car into a big plate glass window, do you think I could keep my windshield intact? Should I wear my seatbelt or not?

My friend has a weed plant in his yard. Could I take a cutting with my teeth and jam it into some compost and get a clone? I don't have any scissors or potting soil.

Quote:

bakedbeings said:
faht does something like what youre describing




Seriously, OP, just follow this link and read up on what faht has played around with, and the meticulously detailed results therein. The vast majority of us have had no reason to play Russian Roulette.


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OfflineBeefSupremeJr
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Re: Mushroom to sterile grain to spawn? [Re: B Traven] * 2
    #28152828 - 01/22/23 08:37 PM (1 year, 2 months ago)

OP Just clone the fruit to agar.  typically the plates gonna contam and youll have to work to get it cleaned up on subsequent plates.  once you have a clean culture then go to grain.

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OfflineNoobShroob
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Re: Mushroom to sterile grain to spawn? [Re: BeefSupremeJr]
    #28152839 - 01/22/23 08:44 PM (1 year, 2 months ago)

Im going to make agar soon, i just dont know if itll be soon enough the fruits are still fresh. I have the supplies its more of an issue of timing on my pc. Im sure ill make something work its just difficult. Regardless, do you have any goto fruit to agar tek, or something on cloning or plate to plate to get the cleaner version i dont understand that part much.


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OfflineJW123
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28152844 - 01/22/23 08:49 PM (1 year, 2 months ago)

It is kind of old, but this thread should help.
https://www.shroomery.org/forums/showflat.php/Number/18430998#18430998

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OfflineNoobShroob
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Re: Mushroom to sterile grain to spawn? [Re: B Traven] * 1
    #28152845 - 01/22/23 08:50 PM (1 year, 2 months ago)

Quote:

B Traven said:
If I run my car into a big plate glass window, do you think I could keep my windshield intact? Should I wear my seatbelt or not?

My friend has a weed plant in his yard. Could I take a cutting with my teeth and jam it into some compost and get a clone? I don't have any scissors or potting soil.





see but this is different because i have everything, syringe, pc, fruiting bodies, its not like i dont have "any scissors or potting soil."

i have a sterilized bag already.

even if i got the agar (which is seemingly the big guarantee on success, although not only way) i still dont understand how i can get it into the bag without risk of contam, that why i assumed that going the LC route would seemingly be just as risky possibly less as i wouldnt be opening the entire bag and attempting to reseal it, although i could do this.


Quote:

JW123 said:
It is kind of old, but this thread should help.
https://www.shroomery.org/forums/showflat.php/Number/18430998#18430998




thank you!

also what about just a spore syringe over lc?


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Edited by NoobShroob (01/22/23 08:51 PM)

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OfflineBeefSupremeJr
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28152846 - 01/22/23 08:50 PM (1 year, 2 months ago)

You bet!

in my journal

youll find several writeups regarding agar.  see stro's writeup on cleaning and isolating.

cloning aint no thang.  you basically just split a stipe open and take a little tissue from inside and put it on a plate.  ive done it and had perfectly clean results.

read my friend.  it aint even that much reading.  this shits actually easier done than said.

Edited by BeefSupremeJr (01/22/23 08:55 PM)

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OfflineJW123
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28152850 - 01/22/23 08:54 PM (1 year, 2 months ago)

No problem. As Beef said it really isn't very hard. Keep backups and do a lot of SAB work and it gets easier.

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OfflineB Traven
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28152857 - 01/22/23 08:57 PM (1 year, 2 months ago)

I just use plate pins, ie I let agar plates colonize completely and then wait for them to produce tiny immature fruits on the surface. Since those are forming in a hopefully sterile vessel, I can just pick them up with a flamed scalpel in the SAB and drop them onto new plates. No real need to worry about contamination on the exterior of those pins.

Any fruit that grew in open air is presumed to be covered in contamination, so the goal would be to take a tissue sample from the interior. I think a lot of people basically just pull the fruit apart to access the unexposed interior tissue. Cutting into it is another option, but I think that can create the risk that you're picking up stuff from the surface and pushing it into the sample you're trying to take.

I would think that, for the purposes of taking clones from interior tissue, fresh fruits would keep pretty well for a while in the refrigerator. Maybe long enough for you to get the PC situation worked out.

I wonder if you could take an interior tissue sample and transfer it to an oven-sterilized pyrex container, and then just leave that in the fridge (or wherever). Even if it dries out, it should stay sterile, and you could then try reviving it on agar. If it's  a short enough interval, though, it probably wouldn't even dry out much.


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OfflineBeefSupremeJr
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Re: Mushroom to sterile grain to spawn? [Re: B Traven] * 2
    #28152861 - 01/22/23 09:03 PM (1 year, 2 months ago)

yeah never cut into a fruit.  use your fingers and split it in half and curt from just below the cap in the stipe.

im thinkin if you took a fruit, washed it, put it in ziplock, itd be just fine for quite some time..

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OfflineB Traven
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28152864 - 01/22/23 09:05 PM (1 year, 2 months ago)

Quote:

NoobShroob said:
Quote:

B Traven said:
If I run my car into a big plate glass window, do you think I could keep my windshield intact? Should I wear my seatbelt or not?

My friend has a weed plant in his yard. Could I take a cutting with my teeth and jam it into some compost and get a clone? I don't have any scissors or potting soil.





see but this is different because i have everything, syringe, pc, fruiting bodies, its not like i dont have "any scissors or potting soil."

i have a sterilized bag already.

even if i got the agar (which is seemingly the big guarantee on success, although not only way) i still dont understand how i can get it into the bag without risk of contam, that why i assumed that going the LC route would seemingly be just as risky possibly less as i wouldnt be opening the entire bag and attempting to reseal it, although i could do




My point was just that you're trying to cut out critical steps, and the reason seemed to be that you weren't set up to do certain things yet. You're either ready to do all the steps or you're not. Having what you need for the first and last steps doesn't count.

Yeah, using the port is probably the best way to inoculate that bag. You don't have to use LC, though. You can take an agar plate you're ready to send and make liquid inoculant with it- same idea, but you're not trying to grow mycelium out in the liquid medium. You're basically just sucking myc into a syringe of sterile water and using it to inoculate.

If you get that far, definitely keep some plates in refrigerated storage, too.


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OfflineBeefSupremeJr
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Re: Mushroom to sterile grain to spawn? [Re: B Traven] * 1
    #28152871 - 01/22/23 09:09 PM (1 year, 2 months ago)

Hmm... ive heard of people resuscitating dry myc... another words you could dry the fruit and still clone it.

i think?

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OfflineNoobShroob
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Re: Mushroom to sterile grain to spawn? [Re: B Traven]
    #28152876 - 01/22/23 09:13 PM (1 year, 2 months ago)

i understand, thats real


Quote:

B Traven said:
Yeah, using the port is probably the best way to inoculate that bag. You don't have to use LC, though. You can take an agar plate you're ready to send and make liquid inoculant with it- same idea, but you're not trying to grow mycelium out in the liquid medium. You're basically just sucking myc into a syringe of sterile water and using it to inoculate.





i didnt even know that was possible, agar seems to just open the door for everything.


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OfflineBeefSupremeJr
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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob] * 1
    #28152882 - 01/22/23 09:18 PM (1 year, 2 months ago)

this is the way

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Invisiblebakedbeings
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Re: Mushroom to sterile grain to spawn? [Re: BeefSupremeJr] * 1
    #28153326 - 01/23/23 07:34 AM (1 year, 2 months ago)

you can also clone tissue to a BRF puck if your agar hasnt arrived

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Re: Mushroom to sterile grain to spawn? [Re: NoobShroob]
    #28153333 - 01/23/23 07:43 AM (1 year, 2 months ago)

Quote:

NoobShroob said:
Quote:

B Traven said:
If I run my car into a big plate glass window, do you think I could keep my windshield intact? Should I wear my seatbelt or not?

My friend has a weed plant in his yard. Could I take a cutting with my teeth and jam it into some compost and get a clone? I don't have any scissors or potting soil.





see but this is different because i have everything, syringe, pc, fruiting bodies, its not like i dont have "any scissors or potting soil."

i have a sterilized bag already.

even if i got the agar (which is seemingly the big guarantee on success, although not only way) i still dont understand how i can get it into the bag without risk of contam, that why i assumed that going the LC route would seemingly be just as risky possibly less as i wouldnt be opening the entire bag and attempting to reseal it, although i could do this.


Quote:

JW123 said:
It is kind of old, but this thread should help.
https://www.shroomery.org/forums/showflat.php/Number/18430998#18430998




thank you!

also what about just a spore syringe over lc?




It's not the putting it in the bag you have to worry about when using fruit to clone. It's all the bacteria and spores already present on the piece of mushroom you are using. By going to agar first you can clean up the contamination by transferring 1 or more times to clean agar before sending it to your grain.

IMHO if you are to lazy or impatient to wait and do things right then you should just buy your shrooms. Will save you a lot of heartache and expense.

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Offlinehao
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Re: Mushroom to sterile grain to spawn? [Re: RockinRobot]
    #28153350 - 01/23/23 08:00 AM (1 year, 2 months ago)

Quote:

RockinRobot said:
Quote:

NoobShroob said:
Quote:

B Traven said:
If I run my car into a big plate glass window, do you think I could keep my windshield intact? Should I wear my seatbelt or not?

My friend has a weed plant in his yard. Could I take a cutting with my teeth and jam it into some compost and get a clone? I don't have any scissors or potting soil.





see but this is different because i have everything, syringe, pc, fruiting bodies, its not like i dont have "any scissors or potting soil."

i have a sterilized bag already.

even if i got the agar (which is seemingly the big guarantee on success, although not only way) i still dont understand how i can get it into the bag without risk of contam, that why i assumed that going the LC route would seemingly be just as risky possibly less as i wouldnt be opening the entire bag and attempting to reseal it, although i could do this.


Quote:

JW123 said:
It is kind of old, but this thread should help.
https://www.shroomery.org/forums/showflat.php/Number/18430998#18430998




thank you!

also what about just a spore syringe over lc?




It's not the putting it in the bag you have to worry about when using fruit to clone. It's all the bacteria and spores already present on the piece of mushroom you are using. By going to agar first you can clean up the contamination by transferring 1 or more times to clean agar before sending it to your grain.

IMHO if you are to lazy or impatient to wait and do things right then you should just buy your shrooms. Will save you a lot of heartache and expense.





Exactly

It's almost impossible to have a perfectly sterile fruit, that mean putting bacteria and contam to your future bag...

Afar would be perfect to clean before.

Good luck
Love all

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Invisiblebakedbeings
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Re: Mushroom to sterile grain to spawn? [Re: hao]
    #28153568 - 01/23/23 10:25 AM (1 year, 2 months ago)

not to mention a mail order grain bag is probably bacterial anyway

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Offlinehao
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Re: Mushroom to sterile grain to spawn? [Re: bakedbeings]
    #28153704 - 01/23/23 11:53 AM (1 year, 2 months ago)

The problem is you never know when it was sterilized but the colonizing success depend on that..

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OfflineGreg
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Re: Mushroom to sterile grain to spawn? [Re: hao]
    #28153711 - 01/23/23 11:56 AM (1 year, 2 months ago)

Having worked in a shipping facility as a loader / sorter I can attest that there's a very good chance sterile grain bags get fucked in shipping too.
Some people would literally drop kick and overhand throw packages into the trucks.

You're also trusting whoever the dork is who preps the grains at your chosen vendor to hydrate them properly, pc long enough, and impulse seal them well.

Edited by Greg (01/23/23 11:58 AM)

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OfflineScrewup
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Re: Mushroom to sterile grain to spawn? [Re: Greg]
    #28153720 - 01/23/23 12:00 PM (1 year, 2 months ago)

Lol can confirm. Let alone packages moving during shipping or just dropping from the auto sorter? Let alone the underpaid people who dgaf about your package and will literally go out of their way to drop kick your clearly marked fragile product?


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