Cloning Psilocybin

[canndo]

Ok So keep it simple.
Instead of jars, couldn't I use one foil covered bowl in a pressure cooker?:

-Move the cooker and bowl inside the sterilized environment.
-Use an airtight Rubbermaid (Or wtv you're final grow will sit in)
-Add sterilized substrate from bowl to final container
-Inoculate all over the container
-Add your water shield
-Put on the container cover, wrap in a dark bag, store in a dark warm place

And that way I wouldn't have to take the substrate out of jars...
And there was a mention of Agar, which I read is a red algae derivative. I also read the red Algae is beneficial for healthy growth of many mushrooms.
But the "Kanten" food grade agar available in Oriental stores would work just as well?

You could use a foil covered bowl but that bowl will not be air tight and contamination may get in. Furthermore, the wider your opening, the more chance of contamination. I don't think you understand the process. the correct process care for your myceilum - multiplying it in volume while going from a more to less sterile state. The mushroom when it has fully collonized a substrate is very contaminant resistant. You can handle fully colonized grain with dirty hands (not recommended though), and still be able to spawn your substrate - which no longer needs to be sterile (or, sterile except for you chosen organism - I always marveled at ads for spores that were "viable and sterile").

So I don't see why you are trying to go the way you are - there is no advantage and several dissadvantages to innoculation in place. Taking the substrate out of the jars after it has been colonized is not a problem.

food grade agar or agar agar is perfectly useable but it is more trouble than conventional agar combinations. Why not simply purchase half a pound of pre-formulated agar+cornsteep+ yeast+malt extract or potato dextrose? It disolves readily in water whereas the agar needs to be cut or ground and then cooked - in most of the food style presentations anyway. Remember that agar is only the gelling component, nutrient still needs to be added.

 

[polyarcturus]

i would recommend getting spores(to start) they are pretty cheap, use a greendot credit card and have them sent to a friends house.

cloning from a dry mushroom is difficult you will have to find a way to rehydrate the mushroom before you can even attempt to clone it, honestly ive never attempted it but i think it could be done with enough trys... but that might cost you more in the long run

 

[canndo]

i would recommend getting spores(to start) they are pretty cheap, use a greendot credit card and have them sent to a friends house.

cloning from a dry mushroom is difficult you will have to find a way to rehydrate the mushroom before you can even attempt to clone it, honestly ive never attempted it but i think it could be done with enough trys... but that might cost you more in the long run

Attempting to grow from a deydrated fruit is something for somene who has lots of experience with such things and I blelieve will only serve to frustrate others. As I said, I have tried it with absolutely no sucess - and except for revitalizing some odd strains, see little point in it as spores are so easy to come by - and given that fact that the life cycle of the mushroom is spores - myelium - fruit - spores. It is always best to conform to the growth cycle of the organism we are working to understand.

 

[Alembic]

I have tried, and succeeded... Twice.

Most will tell you this is stupid, but like I said, I did it, twice.

Take your dried mushrooms (if they're not cracker dry, don't expect much success) and dunk them into 3% H2O2 solution. for about 2-3 minutes. Transfer to agar. Wait. I have two dishes with cultivated mycellium in my closet from dried mushrooms.

 

[SnakeByte]

water shelid?

Ya you know... Cuz sometimes the container MAY not always be able to stay at 100% humidity, so you need to spray it down. But direct water contact will harm your culture so you put like a plexiglass on a slant inside with the culture. Then you spray the glass so as not to harm them and keeping the humidity up.

You could use a foil covered bowl but that bowl will not be air tight and contamination may get in. Furthermore, the wider your opening, the more chance of contamination.

I see the logic in that. Though, from everything I know (which seems less and less) what's normally used are preservative jars with two holes in the lids covered by foil.
I mean, neither are really air tight - and while yes the bowl does have a MUCH larger opening, will that actually make a Huge difference?
When in both scenarios the substrate is take directly from the cooker, to a sterilized environment, inoculated, Then packed and put away.
I'll also be using a "Still Air/Glove/Inoculation" Box inside the sterilized environment. It'll be big enough to fit the cooker right inside and still have room for my other tools and to work comfortably.

And that H2O2 Solution idea does not sound too crazy. As it's been mentioned, mushrooms take in O2 l[SUB][/SUB]ike we do correct? And H2O2 releases O2 into water and air.

 

[Alembic]

I've been basically told what I did was stupid a multitude of times for using H2O2, but hey, I made it work.

 

[polyarcturus]

so you plant on inoculating the bowl full of substrate covering with foil then putting that into a high humidity enviroment? why? the foil creates a micro enviroment inside the bowl that will already be very high in humidity, then when you remove the foil i could see an addition of a drip shield(something i need in my setup cause im constantly wiping the lid.. that or more fae, probably more fae...


but considering that you can be careful with you spray and only spary the walls you shouldnt need a drip shield.


and i think you missing the point when i say "cloning" is a dirty process. lots of times contams are grown into the fruit as the cell grow, at this point it is far too late for it to do any damage to the fruits(because they grow so fast consuming the contam) but there can be contams within the fruit and not just the outside, doesnt matter if you got a flow hood or a glove box.

(or open air, i did a clone open air into pcorn recently honestly i expect it to be contamed or not grow)

 

[SnakeByte]

Yeah, exactly. With a shield - just tap the top of the chamber before you open and all the water will fall on the glass, With no real need to be careful of where you spray
I've always thought jars are a pain, especially if you want to just grow some healthy mycelium and not fruit bodies.
I'm trying to keep the steps intact because I know it's a sensitive process but they do make tiny mason jars which would be useful here.
But let's say this is done straight from bought syringe now: I'm just wondering why not skip one step and inoculate the fruiting chamber itself, instead of letting the mycelium form in a jar first?
As canndo said (I paraphrase), that it is only the inoculation and initial birthing that is important.
I don't know If I'm explaining it right, but in my mind it's two birds with one stone.

 

[polyarcturus]

yeah but in a jar you can guarantee its contam free, proper growth, identify special ones. by all mean go ahead and innoc a bulk with spores(people do it all the time) and wait and hope for the best. with jars you can see whats going on and you can time thing better, and save a grow if needed. as far as time consumption i would say inoculating a bulk from the start vs. inoculation with jars would be maybe a week faster, but there is no way to check on growth conveniently, and opening it early could be you grows downfall.

 

[canndo]

I have tried, and succeeded... Twice.

Most will tell you this is stupid, but like I said, I did it, twice.

Take your dried mushrooms (if they're not cracker dry, don't expect much success) and dunk them into 3% H2O2 solution. for about 2-3 minutes. Transfer to agar. Wait. I have two dishes with cultivated mycellium in my closet from dried mushrooms.

Very cool. How long were they dry? (I don't know if it makes a difference, it shouldn't). the H202 is for sterility? or is there some other function. Cap or stem? or sclerotia? And your agar composition? Corn steep? pda? or mea?

 

[canndo]

Ya you know... Cuz sometimes the container MAY not always be able to stay at 100% humidity, so you need to spray it down. But direct water contact will harm your culture so you put like a plexiglass on a slant inside with the culture. Then you spray the glass so as not to harm them and keeping the humidity up.



I see the logic in that. Though, from everything I know (which seems less and less) what's normally used are preservative jars with two holes in the lids covered by foil.
I mean, neither are really air tight - and while yes the bowl does have a MUCH larger opening, will that actually make a Huge difference?
When in both scenarios the substrate is take directly from the cooker, to a sterilized environment, inoculated, Then packed and put away.
I'll also be using a "Still Air/Glove/Inoculation" Box inside the sterilized environment. It'll be big enough to fit the cooker right inside and still have room for my other tools and to work comfortably.

And that H2O2 Solution idea does not sound too crazy. As it's been mentioned, mushrooms take in O2 like we do correct? And H2O2 releases O2 into water and air.

What is "normaly used" are mason jars either sealed or with a hole or two and fiber micro filters between the top lid and the jar. If they have holes in them it is because the intent is to innoculate the substrate in the jar. If this is the case, it is unlikely that anyone would want to empty sterile substrate into another container - that would defeat the entire process. The size of the opening does make a difference depending of course on your environment. I don't like big open containers when I work. Testtime doesn't like ANY open - contaminant catching spaces in his work. I am confident enough and willing to chuck up to 20 percent of my work where I will rarely use filters except when I know that I will be growing mycelium in the jar for more than two weeks.

I do'nt get your "packed and put away" part. Packed? how? Again I think you are mistaking a step. Spawn is fully colonized before it is removed from it's original container.


I know of others who put their cookers in their glove boxes. I think that requires a pretty large glove box - especially if you are opening the lid in the box. I find it better to put alcohol soaked rags around the PC port - remember that this is a pressure cooker and not an autoclave. When the cooker cools, it will begin to draw air back into the body of the cooker. This air will bring contamination in with it. You can put your cooker in your box when it is hot to reduce this effect, or you can set up a rudamentary filter as I do. I then take my jars or bags or whathave you out of the pc while still as hot as I can touch and lay them in my sterile space. I have the advantage of using a true hood so I don't worry too much about such things but I worked and still often work without it.


I would advise you to concentrate on your tecnique and not your environment. There is only so much you can do about your environment, you can refine your tecnique to a great degree, if you get good at it you can do open air transfers and innoculations. It is all in planning your hand movements, your placement, having a set of movement skills that has your suceptable materials exposed for the shortest amount of time. Remember that what causes contamination is most often physical contact with something that is intended for your growth. Touching a bit of mushroom with a contaminated scalpel, touching your agar with a contaminated instrument - that sort of thing. Be clean, be methodical, be reapeatable so that if you fail, you can deduce the cause by going through your steps and discovering your error. But beyond a certain point - don't go nuts with the sterility, especially after your innoculations and initial colonization.


Always remember that nature is a very dirty place - and yet, mushrooms grow there quite well.

 

[canndo]

I've been basically told what I did was stupid a multitude of times for using H2O2, but hey, I made it work.

If it worked and it is adventageous that it worked, then you weren't doing a stupid thing, were you?

 

[canndo]

Yeah, exactly. With a shield - just tap the top of the chamber before you open and all the water will fall on the glass, With no real need to be careful of where you spray
I've always thought jars are a pain, especially if you want to just grow some healthy mycelium and not fruit bodies.
I'm trying to keep the steps intact because I know it's a sensitive process but they do make tiny mason jars which would be useful here.
But let's say this is done straight from bought syringe now: I'm just wondering why not skip one step and inoculate the fruiting chamber itself, instead of letting the mycelium form in a jar first?
As canndo said (I paraphrase), that it is only the inoculation and initial birthing that is important.
I don't know If I'm explaining it right, but in my mind it's two birds with one stone.

You are making a huge presumption - as poly is trying to say. If you jump directly from innoculation to bulk substrate you lose the ability to make your spawn multipoint. You are also presuming that your initial attempt will be sucessful and it very well may not. I say I am comfortable with a 20 percent loss rate - that means that I lose 2 of 10 jars or bags but I am assured that the spawn I do use is good and I won't waste my time spawning with garbage. Furthermore I happen to enjoy the speed of colonization gotten through the ability to shake my jars or bags. As I have said many times here - I can with a little luck create a nearly infinte amount of spawn in a fraction of the time a PF guy gets his little pucks fully colonized - and that is if the core of those cakes is complete and he can't know that.

Honestly my friend, it seems like you are attempting to re-invent the wheel here and it is not necessary.

 

[polyarcturus]

Remember that what causes contamination is most often physical contact with something that is intended for your growth. Touching a bit of mushroom with a contaminated scalpel, touching your agar with a contaminated instrument - that sort of thing. Be clean, be methodical, be reapeatable so that if you fail, you can deduce the cause by going through your steps and discovering your error. But beyond a certain point - don't go nuts with the sterility, especially after your innoculations and initial colonization.


Always remember that nature is a very dirty place - and yet, mushrooms grow there quite well.

,

just had this happen to me, 2 agar cultures of a GT isolation, one i scraped the agar with the scalpel on accident and it is half contaminated half mycelium, the other jar that i did not touch the agar with is 100% not contaminated.

also note i got my clone to start producing mycelium, and that was open air too! like canndo said its all about you method as well as being sterile.

 

[SnakeByte]

I figure the oxygenation from the H2O2 probably had something to do with it.
Hydrogen Peroxide is found in the atmosphere and falls in the rain. Bit of Info

Yeah, exactly. With a shield - just tap the top of the chamber before you open and all the water will fall on the glass, With no real need to be careful of where you spray
I've always thought jars are a pain, especially if you want to just grow some healthy mycelium and not fruit bodies.
I'm trying to keep the steps intact because I know it's a sensitive process but they do make tiny mason jars which would be useful here.
But let's say this is done straight from bought syringe now: I'm just wondering why not skip one step and inoculate the fruiting chamber itself, instead of letting the mycelium form in a jar first?
As canndo said (I paraphrase), that it is only the inoculation and initial birthing that is important.
I don't know If I'm explaining it right, but in my mind it's two birds with one stone.

You are making a huge presumption - as poly is trying to say. If you jump directly from innoculation to bulk substrate you lose the ability to make your spawn multipoint. You are also presuming that your initial attempt will be successful and it very well may not. I say I am comfortable with a 20 percent loss rate - that means that I lose 2 of 10 jars or bags but I am assured that the spawn I do use is good and I won't waste my time spawning with garbage. Furthermore I happen to enjoy the speed of colonization gotten through the ability to shake my jars or bags. As I have said many times here - I can with a little luck create a nearly infinte amount of spawn in a fraction of the time a PF guy gets his little pucks fully colonized - and that is if the core of those cakes is complete and he can't know that.

Honestly my friend, it seems like you are attempting to re-invent the wheel here and it is not necessary.

These are all very true. The biggest one being that I'm assuming there will be no contam.
I'm most likely thinking too industrial. Like stacking up five 6 Quart Rubbermaids instead of dealing with jars lol
What kinds of stores carry rye seed? Brewer shops and the like I'd assume?

 

[canndo]

Health food and whole food stores sell it. Or you can go to feed lots if you want bulk - watch out - some of it has high spore loads.

 

[Alembic]

canndo, I used H2O2 to attempt to kill anything on the outside of the mushroom. The Mushrooms wre dried for a better part of at LEAST 9 months.

 

[canndo]

canndo, I used H2O2 to attempt to kill anything on the outside of the mushroom. The Mushrooms wre dried for a better part of at LEAST 9 months.

Your n202 was not pure so you were putting a percentage of water back on your dried fruit. I did the very same thing with sclerotia to no avail.

 

[polyarcturus]

I know of others who put their cookers in their glove boxes. I think that requires a pretty large glove box - especially if you are opening the lid in the box. I find it better to put alcohol soaked rags around the PC port - remember that this is a pressure cooker and not an autoclave. When the cooker cools, it will begin to draw air back into the body of the cooker. This air will bring contamination in with it. You can put your cooker in your box when it is hot to reduce this effect, or you can set up a rudamentary filter as I do. I then take my jars or bags or whathave you out of the pc while still as hot as I can touch and lay them in my sterile space. I have the advantage of using a true hood so I don't worry too much about such things but I worked and still often work without it.

i always open the Pc while its still, warm to hot, just when it loses pressure after a few hours then i put the jars under a rag to cool. i rarely leave them in the PC to cool but i have in the past with no problems.

 

[testtime]

i always open the Pc while its still, warm to hot, just when it loses pressure after a few hours then i put the jars under a rag to cool. i rarely leave them in the PC to cool but i have in the past with no problems.

Way too lazy and distracted for that. I've let them sit for days before unpacking.