1st shroom grow, need help

T

testtime

Well-Known Member
canndo said:
Poly has gotten the big picture, that it is not in your equipment but in your methods and preparation. Those who rely on anything else will never get the sort of results those who do not.... do.
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Ehh. Religious viewpoint. I can do open air grain to grain and let the jars sit open for 10 minutes while I'm working on it in my ozone chamber.

I do an open air pour of agar to a bunch of dishes at once, let them cool off open, and cap them off.

No contams.

No amount of skill will beat a perfect environment.
 
Brizzy

Brizzy

Active Member
Inoculated today. First hole on first jar I accidentally shot almost a whole cc in one hole...sirenges are touchy.. Ended up using about 5ccs total in the 3 jars. So a little less than 2cc average. Sterilized my hands, work area, sirenge and worked quickly. After all were pregnated i shook the jars a little. My jars are now sitting in my sock drawer that I never get into. Ill check em in a few days
 
canndo

canndo

Well-Known Member
testtime said:
Ehh. Religious viewpoint. I can do open air grain to grain and let the jars sit open for 10 minutes while I'm working on it in my ozone chamber.

I do an open air pour of agar to a bunch of dishes at once, let them cool off open, and cap them off.

No contams.

No amount of skill will beat a perfect environment.
Click to expand...

There is no perfect environment. It is one thing to work in a household or lab that is freshly clean and has a low particulate count but this is a temporary thing and eventually the count will rise, when that happens as it always does, one has to depend upon one's ability to work in such an environment.


Even a class one clean room can be contaminated upstream spreading spores on everything downwind from the original point of contamination. Given that, the best way to consistantly come up with successful innoculations is to refine one's ability not only to work efficiently but to develop a second sense regarding the unseen.
 
T

testtime

Well-Known Member
canndo said:
There is no perfect environment. It is one thing to work in a household or lab that is freshly clean and has a low particulate count but this is a temporary thing and eventually the count will rise, when that happens as it always does, one has to depend upon one's ability to work in such an environment.


Even a class one clean room can be contaminated upstream spreading spores on everything downwind from the original point of contamination. Given that, the best way to consistantly come up with successful innoculations is to refine one's ability not only to work efficiently but to develop a second sense regarding the unseen.
Click to expand...
I don't want to be harsh, but I was very specific in what makes it perfect, and you are not addressing it.

Ozone. High levels.

When I place things INTO it they BECOME sterile. It has little to no effect on myc, at least during the limited exposure, and when you cap a jar (or petri dish) that contains ozone, the ozone then reacts with the surface for a bit and disappears.

So you are left with a sterile clean environment with no residue.

Sure, I need a BIT of skill during petri isolations, since I am dealing with contams in the very beginning, but those were introduced as part of the "clone a slice of mushroom" process.

But the act of opening the contammed dish starts to sterilize, and anything that releases to the air is destroyed QUICKLY. Imagine opening the dish into an environment that is 99% bleach vapor.

I've been testing. I've been growing open jars of corn that are filled with myc, and nothing else. I've got other open jars that have not been nocced, and they sit there, no growth, no contams.

If it isn't PERFECT, it is unmeasurably close.

Everything else that everyone goes crazy with such as g2g? No issues, and certainly no skill tuning.
 
T

testtime

Well-Known Member
canndo said:
All friendly I assure you, Testime is a pal. Drop into the Politics area if you want disagreement.
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No problems at all. I just think I have something here, and someday if I get canndo (or someone of equivalent high quality, not many of them) to take it seriously, then maybe others might benefit. Which would make me happy.

But until then, I gotta drop it in where it fits. Which was as a response to the statement.

All very friendly. Especially with canndo's motto:

  • Argue for your opinion because it is true, not because it is your opinion.​

Read a bit on this concept and then maybe you might find a use for it inside a glove box/environment FULL TIME.

http://lmgtfy.com/?q=use+of+ozone+to+sterilize+hospital+operating+rooms
 
canndo

canndo

Well-Known Member
testtime said:
No problems at all. I just think I have something here, and someday if I get canndo (or someone of equivalent high quality, not many of them) to take it seriously, then maybe others might benefit. Which would make me happy.

But until then, I gotta drop it in where it fits. Which was as a response to the statement.

All very friendly. Especially with canndo's motto:

  • Argue for your opinion because it is true, not because it is your opinion.​

Read a bit on this concept and then maybe you might find a use for it inside a glove box/environment FULL TIME.

http://lmgtfy.com/?q=use+of+ozone+to+sterilize+hospital+operating+rooms
Click to expand...


A simple test then is in order.


Grow a full dish of trich, another one of micor, place them in your chamber with a small method of air disruption and do your transfers.



If you get to a place where you can do that without contamination on agar, I will change up my box and pump ozone in.
 
T

testtime

Well-Known Member
canndo said:
A simple test then is in order.


Grow a full dish of trich, another one of micor, place them in your chamber with a small method of air disruption and do your transfers.

If you get to a place where you can do that without contamination on agar, I will change up my box and pump ozone in.
Click to expand...
Very cool. Good suggestion.

The ozone generator itself has a fan and it is placed right behind the work space, facing forward.

Is that acceptable?

Am I allowed to open the tric and micor plates for a while BEFORE I open the agar dishes? If so, how long? I'd prefer it to be 60 minutes for that level of concentrated spores. I'd like to give the ozone a chance to fry it, and from your perspective, if the ozone doesn't work, it'll just end up circulating the spores anyway.

And how to I generate micor? I can get tric off the trees outside.
Oh, typo, (google is my friend). You meant mucor.
Again, how do I generate it on demand?
 
C

ChesusRice

Well-Known Member
Brizzy said:
Inoculated today. First hole on first jar I accidentally shot almost a whole cc in one hole...sirenges are touchy.. Ended up using about 5ccs total in the 3 jars. So a little less than 2cc average. Sterilized my hands, work area, sirenge and worked quickly. After all were pregnated i shook the jars a little. My jars are now sitting in my sock drawer that I never get into. Ill check em in a few days
Click to expand...

Make your own spore prints when you fruit. Never have to worry about syringes again
 
Brizzy

Brizzy

Active Member
I'm all for friendly debate. Looks like you guys are actually getting somewhere, rather than throwing out random so called "facts" that I've seen others do on this site..and in real life for that matter
 
C

ChesusRice

Well-Known Member
Brizzy said:
Is that a difficult/expensive process? I assume it takes lots of know how
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Easy
mushroom cap
aluminum foil
sterilized water (try walgreens)

Put cap on aluminum foil
place sterilized jar over cap and alumimun foil
let sit over night
fill syringes up with sterilized water
flow over aluminum foil or make a soup with your spore print and sterilized water (bowl of water with print in it)

spore syringes

Keep it clean though, and the oven door trick works nice
 
canndo

canndo

Well-Known Member
testtime said:
Very cool. Good suggestion.

The ozone generator itself has a fan and it is placed right behind the work space, facing forward.

Is that acceptable?

Am I allowed to open the tric and micor plates for a while BEFORE I open the agar dishes? If so, how long? I'd prefer it to be 60 minutes for that level of concentrated spores. I'd like to give the ozone a chance to fry it, and from your perspective, if the ozone doesn't work, it'll just end up circulating the spores anyway.

And how to I generate micor? I can get tric off the trees outside.
Oh, typo, (google is my friend). You meant mucor.
Again, how do I generate it on demand?
Click to expand...

I got a couple of generators laying around - they were threatening to outlaw them in my state and I figured it would be a good idea to pick some up. Frankly the stuff scares me. I have kept up with most commercial sterilization tequniques through the years and have never come across a strictly ozone environment for sterilization. As we get old we tend to begin to believe we know everything and we have seen everything. What happens is that we "make stuff up" - it is a long theory and everyone is suseptable to it even as they watch themselves do it. They tend to create their own reality by using their "logic" or "common sense". The result is a sort of mental sclerosis. I am as guilty as anyone else and discounted your ozone set up. If you hadn't interjected it in the appropriate place, more than once, i might have continued to discount it, after all I know EVERYTHING there is to know about mushrooms and tissue culture and no one can ever teach me anything.

Let's talk about the particulars - maybe a new thread?


And yes, mucor. thanks.
 
T

testtime

Well-Known Member
canndo said:
As we get old we tend to begin to believe we know everything and we have seen everything. What happens is that we "make stuff up" - it is a long theory and everyone is suseptable to it even as they watch themselves do it. They tend to create their own reality by using their "logic" or "common sense"

.Snip.

Let's talk about the particulars - maybe a new thread?
Click to expand...
When I am observing something that is a result of something I did, I spent a decent amount of time analyzing, rationalizing, and coming up with a story of how it got there. It could be right, it could be wrong, but it is a starting point of research or a discussion to take it to a place it makes sense, rather than "is a story".

I also spend a serious amount of time analyzing my own thought processes. I want to know WHY I think that, and is the starting point a logical or emotional place? That is why religion (any type) has so little space in my head, it does not survive the process. Having kids, and imparting this style of mental process is a challenge, but when they teach their friends, I know I did something right.

Yeah, I'll start a new thread for this alone.
 
canndo

canndo

Well-Known Member
testtime said:
When I am observing something that is a result of something I did, I spent a decent amount of time analyzing, rationalizing, and coming up with a story of how it got there. It could be right, it could be wrong, but it is a starting point of research or a discussion to take it to a place it makes sense, rather than "is a story".

I also spend a serious amount of time analyzing my own thought processes. I want to know WHY I think that, and is the starting point a logical or emotional place? That is why religion (any type) has so little space in my head, it does not survive the process. Having kids, and imparting this style of mental process is a challenge, but when they teach their friends, I know I did something right.

Yeah, I'll start a new thread for this alone.
Click to expand...

You do know I was talking about ME, and not you, right?
 
T

testtime

Well-Known Member
canndo said:
You do know I was talking about ME, and not you, right?
Click to expand...
Yes. And I was talking about me, since of course the universe (and the recently discovered 17 billion earthlike planets in the goldilocks zone) all revolve around me.

It will be a week or so before I dedicate the ozone space to experiments. It is currently filled with projects in process and they need a bit of time. While I am sure my typical usage is sterile, i've never introduced a visible quantity of spores into the environment.

Here is the test that I will be doing, and starting a new thread on then:


Place stack of "clean" numbered agar dishes/jars in ozone area. These do NOT have to be sterilized and can be handled with bare hands while placing in. Leave the bottoms open and the tops stacked in a pile on the side (shaking your head yet?) I already do this with 100% success on dish making and waiting a week before use.

LME/DEX/pinch of yeast extract/agar boiled then poured into quart jars.
Jars are PCed.
Jars are cooled.

Boil if necessary, or if still hot:

Place jar in ozone area.
Wait a few minutes.
Open wide.
Pour over each dish, like SYRUP on pancakes.
Well, close. Try not to splash the edges and use too much.
Cap agar jar if any left over.
Wait for it to cool.
Note: We have not placed the lids on, and this is wide open.
When cool (20-30) minutes, place lids on dishes/jars.

Stack'em up, push them aside, wait a week to prove no contams in dishes to start.

Ok, this was standard procedure for me.

Next:
Create a tric dish. Go outside and find some green crap groing on a tree, place it into an agar disk, and let it sit for a couple of days.
Also a mucor one: Tell me. I ASSUME i can take some old coir/verm sitting in a bucket and wait for it to smell like sweet/sour apple, then take some of that and put it in an agar disk and let it sit for a week or so.

After contam dishes created:

Place stack of agar plates in ozone area.

Open #1, wait 5 minutes, close it (prove the environment)
Open contam plates, open #2, wait 5 minutes, close it.
Close contam plates, #open #3, wait 5 minutes, close it.
Open each of the remaining plates in sequence, waiting some time, and closing them.

This test should prove the plates are good, the initial environment is good, whether or not concentrated spores will be neutralized, and if they are not (immediately), how long until they are.

Did I miss anything?
 
canndo

canndo

Well-Known Member
testtime said:
Yes. And I was talking about me, since of course the universe (and the recently discovered 17 billion earthlike planets in the goldilocks zone) all revolve around me.

It will be a week or so before I dedicate the ozone space to experiments. It is currently filled with projects in process and they need a bit of time. While I am sure my typical usage is sterile, i've never introduced a visible quantity of spores into the environment.

Here is the test that I will be doing, and starting a new thread on then:


Place stack of "clean" numbered agar dishes/jars in ozone area. These do NOT have to be sterilized and can be handled with bare hands while placing in. Leave the bottoms open and the tops stacked in a pile on the side (shaking your head yet?) I already do this with 100% success on dish making and waiting a week before use.

LME/DEX/pinch of yeast extract/agar boiled then poured into quart jars.
Jars are PCed.
Jars are cooled.

Boil if necessary, or if still hot:

Place jar in ozone area.
Wait a few minutes.
Open wide.
Pour over each dish, like SYRUP on pancakes.
Well, close. Try not to splash the edges and use too much.
Cap agar jar if any left over.
Wait for it to cool.
Note: We have not placed the lids on, and this is wide open.
When cool (20-30) minutes, place lids on dishes/jars.

Stack'em up, push them aside, wait a week to prove no contams in dishes to start.

Ok, this was standard procedure for me.

Next:
Create a tric dish. Go outside and find some green crap groing on a tree, place it into an agar disk, and let it sit for a couple of days.
Also a mucor one: Tell me. I ASSUME i can take some old coir/verm sitting in a bucket and wait for it to smell like sweet/sour apple, then take some of that and put it in an agar disk and let it sit for a week or so.

After contam dishes created:

Place stack of agar plates in ozone area.

Open #1, wait 5 minutes, close it (prove the environment)
Open contam plates, open #2, wait 5 minutes, close it.
Close contam plates, #open #3, wait 5 minutes, close it.
Open each of the remaining plates in sequence, waiting some time, and closing them.

This test should prove the plates are good, the initial environment is good, whether or not concentrated spores will be neutralized, and if they are not (immediately), how long until they are.

Did I miss anything?
Click to expand...

I am presuming that your agar is either pda or some other common nutrient base.


Other than that, you will have shown that the two most common contaminations, one fungi imperfecti and one bacterial are nulified through the use of ozone alone.

I believe you have already shown that mycelium is not adversely affected. But we don't know if perhaps the genetics have been compromised in some way. I'd be satisfied however just to see the mycelium remain viable while the other contaminations are rendered incapable of sporeulating/spreading or germinating.


And no - I am quite certain that those planets and the beings that inhabit them revolve around me, quite certain.
 
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