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mycohomme
stranger withcandy
Registered: 02/25/04
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multispore agar innoculation
#2387060 - 02/28/04 04:57 PM (19 years, 10 months ago) |
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Does anyone have any experience, tips, or suggestions for a multispore agar innoculation? I have spore prints, and I have read some of the documents regarding techniques. Any advice though would be greatly appreciated.
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Anonymous
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Re: multispore agar innoculation [Re: mycohomme]
#2387303 - 02/28/04 07:31 PM (19 years, 10 months ago) |
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It helps to hold the syringe needle down to allow the spores to collect at the tip...only one drop in say 3 or 4 well spaced places on the plate. Avoid using large drops - minimize the amount of water on the plate...that's just my 2 cents worth. There was a thread on this just a few days ago...use the search function. Someone with more expertise may have better input....
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llamaboy
the weasel thatsnagged the bee

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Re: multispore agar innoculation [Re: mycohomme]
#2387845 - 02/28/04 11:23 PM (19 years, 10 months ago) |
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since you say you have a spore PRINT...i'd suggest using an innoculation loop...or if you don't have that maybe a needle of some sort...just scrape the print with the loop or the needle...and then drag it across the agar in a sorta zigzaggy pattern. if you don't see any spores on the loop or the needle, don't worry, there are some there...then you can do your whole transfering of best growing mycelium or whatever.
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the man
still masked


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Re: multispore agar innoculation [Re: llamaboy]
#2388044 - 02/29/04 01:25 AM (19 years, 10 months ago) |
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i think it would be very interesting to put various strains of cubies on one agar dish. all on different parts. then see what one grows faster, fluffier, or if it takes over other myc. (if you could tell)
peace
-------------------- And Moses Said "Let my mushrooms grow!"
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llamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
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Re: multispore agar innoculation [Re: the man]
#2388069 - 02/29/04 01:46 AM (19 years, 10 months ago) |
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it may be, but i doubt that's what he means by multispore :P
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the man
still masked


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Re: multispore agar innoculation [Re: llamaboy]
#2388093 - 02/29/04 02:03 AM (19 years, 10 months ago) |
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i know just a thought and doesnt need its own thread
-------------------- And Moses Said "Let my mushrooms grow!"
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Anonymous
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Re: multispore agar innoculation [Re: mycohomme] 1
#2388469 - 02/29/04 08:42 AM (19 years, 10 months ago) |
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My bad...disregard my idiocy. I'm going to learn how to read soon, but I guess I'm hooked on chronic...
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tripndicular
My Minds Eye IsRhizomorphic

Registered: 08/25/02
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Re: multispore agar innoculation [Re: mycohomme] 1
#2388620 - 02/29/04 10:18 AM (19 years, 10 months ago) |
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When you first streak a petri filled with agar ...it is a multi spore germination at first , you in time will harvest particular strains by "issolating" the best myc produced i.e. fastest , most rhizo development , etc .... You are trying to grow myc that does not sector , meaning two different types of myc Get copy of TMC , best investment you could make . Good Luck
-------------------- Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will . So with that said here is our mission statement . Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !
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Blue Helix
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Re: multispore agar innoculation [Re: mycohomme] 1
#2389672 - 02/29/04 09:49 PM (19 years, 10 months ago) |
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I have TMC and GGMM. Forget about sectoring with cubes because you'll just be wasting your time. Cubes ALL grow exceptionally well. I have NEVER seen a germination that didn't support vigorous fruiting. Maybe if we were growing some hard-to-fruit species of mushroom, it's make sense to sector, but with cubes anything that isn't contaminated will produce TONS of mushrooms if you grow them out. Another thing all those "cottony" strains that appear on agar--and ALL multi-culture spores innoculations start off that way--will turn rhizomorphic given a good growing environment. I challenge anyone to show me a cube culture that is not contaminated that doesn't fruit like gangbusters. So stop worrying about cottny stuff, sectoring, and all the crap. That's stuff you need to worry about if you are growing something else, not cubes. Anyway, if you have a decent spore print--and forget about spore solutions with agar--you have two options that will prove successful when innculating an agar surface: 1) Work faster 2) Work cleaner Within reason you need both, but you can substitute a little speed for a HEPA filter room. Likewise, you can substitue a little cleanliness if you can innoculate real fast, say the surface is exposed for less than 5 seconds. And as for the innoculation loop, it's fine, but in my experience it just isn't practical to use it fast. You end up spending too much time dinking around with it, and touching the agar surface never works better than just getting the spores to fall on the surface because you end up damaging the surface. Here is how to do it: First, you should be wearing latex or, better yet, those rubber or neoprene dish washing gloves, and the gloves should have been rinsed on the outside with rubbing alcohol or denatured ethanol or the like. Also, keep in mind that you should NEVER breath onto the agar surface. You should never have your hair or bare skin over the exposed agar surface either at any point--remember your whole body is a shower of contamination, even if you are a clean person. Now get a good metal scrapy thing. A good set of tweesers works pretty well. Just heat the scrapy thing up in a blow torch flame. Wait a minute or so for it to cool. And then in one smooth motion, take a hand with the spore print paper between your index and thumb and open the jar or petri with the ball that hand. During the entire process, you should not lay down the lid or face it upwards. Just keep it clenched in the hand that has the spore pring between the index finger and thumb. Practice this beforehand if you have to. Be smooth. Turning the spore print sideways or facing downward on your index finger above the agar, take the tweesers or whatever in you other hand and scrape it with the scrapy thing until the tiniest bit of spores fall on the agar. If you can see a speck, you have PLENTY. Under no circumstances touch the agar surface with anything, including the scrapy thing. Rapidly close the jar or petri dish. It's my experience that you want to do all that in less than 10 seconds and in a draft-free room and a sterile hard surface. Remember be smooth and work fast and you won't need even a HEPA filter.
Edited by Blue Helix (02/29/04 09:56 PM)
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mycohomme
stranger withcandy
Registered: 02/25/04
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Re: multispore agar innoculation [Re: Blue Helix]
#2391932 - 03/01/04 04:48 PM (19 years, 10 months ago) |
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thanks for all the replies! I read though that you should rehydrate the spores in h2o. is this not necessary?
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Anonymous
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Re: multispore agar innoculation [Re: Blue Helix]
#2392151 - 03/01/04 06:07 PM (19 years, 10 months ago) |
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The site says we are trying to stop misinformation.
Please reread the books you claim to possess. Becasue I don't think you understand them.
Cubensis is not unique. It to contains good dikaryons and bad dikaryons, and isolation of individual dikaryons and testing them at different environmnetal parameters will give you a REAL understanding of this.
If you wan't to state that itis not neccesary to get good fruitings, that is one thing, but to imply that cubensis doesn't obey the same rules as every other mushroom with the same reproduction system, or every other living thing on the planet is just stupid.
Inoculation loop works great. You can inoculate agar with spore solutions. And scraping a print directly onto agar is not the cleanest and safest method of getting spores from print to agar. A loop is far safer, and more controlled.
Not all cottony sectors will become rhizo. It is very possible to isolate non fruiting sectors from a very diluted multispore inoculation.
I think your heart is in the right place, but you really are simplifying things way to much.
If you wan't to see the full variety within each Strain, you got to do what you consider a waste of time.
Not all Dikaryons are created equal in each and every environment. Nature doesn't remain 80 F, 95 % humidity all year long. The seasons change, and life changes with it.
Just like there are short people, tall people, skinny people, fat people, smart people, dumb people, etc........... There are differences between Strains , and individual dikaryons of cubensis.
If you haven't seen a bad cubensis fruiting, you haven't really seen a good one yet either.
JUST MY TWO CENTS.
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Blue Helix
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Re: multispore agar innoculation [Re: ]
#2392420 - 03/01/04 07:47 PM (19 years, 10 months ago) |
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Could be, could be... I am just saying I have never seen a cubensis strain that did not show rhizomorphic character before fruiting in the casing layer, and I have never came across a cubensis strain that did not fruit. I'd say my experience is the norm too considering there are thousands of folks around who are using 1/2 pint grain jars as their petri dish in a little tek called PF Tek. Of the many thousands of jars, each like a petri dish, I can't remember the last time I read about a cake that did not fruit unless contamination took down the cake. Doesn't that seem a little odd to you? Thousands upon thousands of jars and they all seem to fruit well. I think the real challenge would be to find a strain that didn't fruit well. That might be about as uncommon as an albino mushroom or the like, one in tens of thousands probably.
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@cro
new name


Registered: 12/07/02
Posts: 1,224
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Re: multispore agar innoculation [Re: Blue Helix]
#2392499 - 03/01/04 08:16 PM (19 years, 10 months ago) |
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Quote:
Blue Helix said: Of the many thousands of jars, each like a petri dish, I can't remember the last time I read about a cake that did not fruit unless contamination took down the cake. Doesn't that seem a little odd to you? Thousands upon thousands of jars and they all seem to fruit well.
Doesn't seem strange at all to me. When you're referring to multispored inoculations in a jar you have many different substrains in that jar, so if there was a substrain which didn't fruit it would just be shadowed by the other substrains which do. But that leads to the rest of the question, you now have part of a jar completely useless for your purposes. If you were to have isolated a substrain which is known to be productive you wouldn't have that substrate going to waste. Plus the benefit of any other characteristics you isolated it for in the first place.
Peace - @cro
--------------------
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Blue Helix
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Re: multispore agar innoculation [Re: @cro]
#2392728 - 03/01/04 09:10 PM (19 years, 10 months ago) |
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I personally don't use jars--way too hard compared with agar and mycellium syringes--but I don't believe that a significant part of the cake is warring strains. I'd bet all of that fighting is done on a very small scale. I have no proof, but that's what I'm betting.
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Psilygirl
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Re: multispore agar innoculation [Re: Blue Helix]
#2392901 - 03/01/04 09:58 PM (19 years, 10 months ago) |
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You dont use jars?? So you go straight from agar to a casing???
-------------------- "Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows." Puget Sound Mycological Society
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Blue Helix
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Re: multispore agar innoculation [Re: Psilygirl]
#2393083 - 03/01/04 11:13 PM (19 years, 10 months ago) |
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Ooops, I do use jars. What I meant is that I don't use spore-injected jars. I use a 1/2 pint agar jar, let it grow out, break up the surface with a scrapy thing, pour in 20 ml or so of h2o2 on the broken pieces, shake up the jar, pour in a couple hundred ml of sterile water, shake up the jar, and then I use the solution to inject birdseed/brown rice/vermiculite jars using a really big syring (60 ml). When those are done colonizing--I give them about 7 days to colonize and another 4 days just for good measure--I apply the casing, incubate the tray for 5 more days covered with syran and in the dark, and eventually fruit out the tray. Fruiting chamber is misted using an ultrasonic on a timer and kept circulating using a cool mist and a PC fan (both on an inverted timer of the ultrasonic).
Edited by Blue Helix (03/01/04 11:14 PM)
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Psilygirl
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Re: multispore agar innoculation [Re: Blue Helix] 1
#2393122 - 03/01/04 11:22 PM (19 years, 10 months ago) |
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gotcha. just confused on your wording.
good luck
-------------------- "Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows." Puget Sound Mycological Society
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ragadinks
MrBeatle


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Re: multispore agar innoculation [Re: Blue Helix]
#2394458 - 03/02/04 09:23 AM (19 years, 10 months ago) |
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How do you prepare and pour in the H2o2 and sterile water into the agar jars ? Do you use a glovebox ?
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Psilygirl
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Re: multispore agar innoculation [Re: ragadinks]
#2395220 - 03/02/04 01:25 PM (19 years, 10 months ago) |
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i'm not sure a what point you're asking about, just make sure you're not adding H2O2 to anything that doesn't have mycelium
-------------------- "Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows." Puget Sound Mycological Society
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ragadinks
MrBeatle


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Re: multispore agar innoculation [Re: Psilygirl]
#2395627 - 03/02/04 02:53 PM (19 years, 10 months ago) |
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I just wonder if I can pour the water into the jar without risking to contaminate the jar again.
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Psilygirl
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Re: multispore agar innoculation [Re: ragadinks]
#2395759 - 03/02/04 03:25 PM (19 years, 10 months ago) |
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as long as you sterilize the water, the water itself wont bring in any contams. however, whenever you open up the lid to the jar you of course risk bringing in new contams. this is where a flowhood/glovebox may help reduce airborne contams getting in while the jar lid is open while you pour in the water.
-------------------- "Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows." Puget Sound Mycological Society
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ragadinks
MrBeatle


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Re: multispore agar innoculation [Re: Psilygirl]
#2395834 - 03/02/04 03:42 PM (19 years, 10 months ago) |
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I have got some bottles with a self healing lid and I wonder if I could probably get the water inside with the help of a syringe under totally unsterile conditions ?
That is what the bottles look like:
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mycohomme
stranger withcandy
Registered: 02/25/04
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Re: multispore agar innoculation [Re: Psilygirl]
#2395844 - 03/02/04 03:44 PM (19 years, 10 months ago) |
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i still dont think i've heard anything regarding the necessity (ot not) of rehydating the spores before going to agar? Do you all just use an innoc. loop and transfer a bit of spores to the agar? or do you maybe make up a syringe, and add a small amount to the agar?
how many transfers does it generally take to get a pure strain?
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mycohomme
stranger withcandy
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Re: multispore agar innoculation [Re: ragadinks]
#2395880 - 03/02/04 03:52 PM (19 years, 10 months ago) |
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raga,
I'm assuming you meant "sterile conditions" not "unsterile" it seems they would do the trick. however, you cant ever be totally sterile. you have to flame the needle to redhottedness, and let it cool of course, so as not to damage those bottles. The needle could still have some contaminants. I dont think its very possible to be completely sterile, if you read one of the previous posts, they something about working cleaner and working faster.
And there is a point where you are trying to hard, you want to do the best you can to keep things sterile, but you can never be perfect. So it just depends on what its worth to you.
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@cro
new name


Registered: 12/07/02
Posts: 1,224
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Re: multispore agar innoculation [Re: ragadinks]
#2395946 - 03/02/04 04:06 PM (19 years, 10 months ago) |
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The only problem I would see would be doctors are usually only pulling out a couple cc's, not much of a problem. You'd be pulling out at 10-12 cc's which would create a vacuum, I'm not sure if it would let you pull out that much. I suppose you could suck air in to the syringe while it's in the presence of a flame then just shoot that into the jar and suck up the liquid afterwards, of course you would have to do this in a couple steps otherwise you'd still be creating a pressure gradient just in the opposite direction.
Peace - @cro
--------------------
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Anonymous
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Re: multispore agar innoculation [Re: Blue Helix]
#2396022 - 03/02/04 04:27 PM (19 years, 10 months ago) |
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There is a very big difference between rhizo in a casing layer, and rhizo on the substrate.
Yes multispore inoculations of substrate guarentees, minus contamination, that you get a fruiting strain. The question is is it the best you can get from any given print. Unless it is by chance, no.
Every population looks like a bell curve, you have the majority right in the middle, you have some that are bad, and some that are above average. If you isolate and test alot of dikaryons, you can find those above average isolates, without rolling the dice.
Is it practical for those just trying to get a fruiting strain, NO. Is it a waste of time for those who want to do the work, NO.
I have never isolated and tried to fruit a non-fruiter either. By selecting vigourous growth, I avoid this. But I have isolated dikaryons from a single print, that showed variability. Bad fruiters, good frutiers, and great fruiters. To me it is worth the effort.
I have never fruited a single PF TEK Jar. I never even tried.
If your are looking for ease and good results. Multispore Jars of substrate, and fruit them.
If you are looking to select something nice, isolation of individual dikaryons is a must, and even better is isolating and breeding monokaryons.
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Blue Helix
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Re: multispore agar innoculation [Re: ragadinks]
#2396068 - 03/02/04 04:42 PM (19 years, 10 months ago) |
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The point of using the H2O2 is so that I don't need a glovebox. Any spores that get in the H2O2 water will be killed. Also by rinsing the agar pieces with pure H2O2, you assure they are clean too, including the sides of the jar which could carry contamination even if the surface does not. I never use a glovebox, but I'm not a slob either. Once you have mycellium growing, you can use H2O2 to do just about anything without need for a glovebox. And contrary to popular belief, H2O2 doesn't kill mycellium very well. It doesn't even stun it in the setting of a mycellium syringe. How do I know? My jars are about 75% complete in 4 days usually. They typically are done is 7. If that's stunned mycellium, then I don't care; it's fast enough for me.
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Blue Helix
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Re: multispore agar innoculation [Re: ]
#2396086 - 03/02/04 04:47 PM (19 years, 10 months ago) |
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When one gets about 11 grams per cake, does it really matter? In my experience, most germinations can yield around 10-12 grams per cake in the first three flushes given proper conditions, yet I often hear of much lower fruitings. I often see trays shown around here that look to be in pretty bad shape, even though they are fruiting. The point is, I think trying to isolate a strain that yields that tiny bit extra is really a moot point when it looks like the biggest concern for me, and probably others, has nothing to do with the strain. Just getting the conditions and timings of the whole procedure right is the real trick in this whole game, not the strain. When you can say that EVERY tray you are laying is getting 10+ rock hard dried (as in they snap like a cracker) grams per cake, then we can talk about strains. Until then I think we're talking above our heads here.
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ragadinks
MrBeatle


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Re: multispore agar innoculation [Re: @cro]
#2396210 - 03/02/04 05:32 PM (19 years, 10 months ago) |
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You are right with the vacuum. But when they give an infusion to someone they must have some special thing that let's (sterile?) air into the bottle so that no vacuum is created in there. I have to ask a doc about that ...
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ragadinks
MrBeatle


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Re: multispore agar innoculation [Re: mycohomme]
#2396222 - 03/02/04 05:36 PM (19 years, 10 months ago) |
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Quote:
I'm assuming you meant "sterile conditions" not "unsterile"
Sorry, I meant "sterile conditions". But I think it is important to be as sterile as possible. Especially when you are living in an very unsterile environment as I do
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Anonymous
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Re: multispore agar innoculation [Re: Blue Helix]
#2396254 - 03/02/04 05:44 PM (19 years, 10 months ago) |
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Well I have higher standards then you, and I actually enjoy looking at a larger sample of the populations of each strain.
My yields are consistently higher then those you are stating, so I guess I am allowed to talk Isolation. Thanks for your permission.
Shrooms don't get rock hard. Not even sclerotia gets rock hard.
The EASIEST WAY TO IMPROVE YIELDS in a stable environmnet is Isolation and testing of MANY INDIVIDUALS within a population.
Yes it is EVERY TRAY, because I am fruiting single dikaryons not rolling the dice and settling for PAR. I play for birdies in golf, and I look for ABOVE AVERAGE YIELD in cubensis.
So back to the original point.
Yes you can multispore on agar, isolate multiple individuals from a single petri dish, and colonize substrate with a single dikaryon, and keep cultures of the best one you find for your particular environmnet. You can do so in as little as one transfer from the original multispore plate, or as many as #, depending on the concentration of spores you inoculate with, and the amount that germinate, and the size of the transfer wedge. You will also be able to determine the cleanliness of the spores before inoculating jars of substrate. You will get a real appreciation for the quantity of individuals within a single print from a single strain, and there variability.
If you just want shrooooms dude, then use a syringe and shoot up pftek jars, and get your doses.
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Blue Helix
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Re: multispore agar innoculation [Re: ]
#2396402 - 03/02/04 06:26 PM (19 years, 10 months ago) |
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Teonan, I don't have a lab. In my opinion, the kind of work you are talking about requires a HEPA filtered lab and thousands of dollars worth of equipment to maintain a very precise environment and thus eliminate other variables that would otherwise grossly influence the outcome. Maybe in your estimation spending that kind of money and setting up such a lab is "the easiest way to improve yields" but in mine, it's not. Aside from that, I'd be curious what sort of yield increase you are getting and all about this variability you say you are finding. The reason I am skeptical of sectoring and isolating strains is because the only folks who seem to beat this drum are those who have a vested interest in selling strains. Folks who sell strains place their entire business viability on fact that their strain is better than most of the ones you'd get by chance. They have a VESTED interest in convincing you of what they are claiming, and indeed, if it's not true, they are selling just a dream. It's a bit like the so-called traits that are reputed to be associated with spores found in various parts of the world. As far as I know, spores don't carry many unique traits, be them B+ or B or Amazonian or whatever, yet the spore vendors swear they are really very different. Are they? I suppose spores are as different as an Asian man is from a white man (not much), but to get an "Arnold Schwarzenegger" mushroom, you'd need a clone of a strain which proved to have super strength. Can you find such a strain by sectoring? Yeah, you can but it's going to take a lot of work. You have to make it to the fruiting stage time and again without other factors interfering with your observations or else you can't say what is related to the strain and what is just dumb luck. Doing that means a lab and money. As for "go do some shrooms dude," the experience tends to be a little on the super intense side for me, even given really modest doses, and as a result I don't do it often. But to each his or her own. I certainly don't feel superior to someone else who gets high on shrooms on a monthly basis or even every weekend.
Edited by Blue Helix (03/02/04 06:31 PM)
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daywalker
Do you dream in color?

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Re: multispore agar innoculation [Re: mycohomme]
#2400749 - 03/03/04 10:46 PM (19 years, 10 months ago) |
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Quote:
mycohomme said: i still dont think i've heard anything regarding the necessity (ot not) of rehydating the spores before going to agar? Do you all just use an innoc. loop and transfer a bit of spores to the agar? or do you maybe make up a syringe, and add a small amount to the agar? ...
I am also waiting for that answer. I hope to be working with some agar in the next week and have a print I'd like to try. Taken it upon myself to acquire the tools so is it "spores-to-agar" or "spores-to-h20-to-agar"? If germination will occur with using a loop to streak the agar - no rehydration, then I am all for it!
-------------------- "I suggest we ... learn to love our ... selves before it ... becomes illegal."
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Psilygirl
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Re: multispore agar innoculation [Re: daywalker]
#2401065 - 03/04/04 12:56 AM (19 years, 10 months ago) |
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no, you dont need to rehydrate it, it rehydrates itself on the agar.
just sterilize an innoc. loop (use isopropyl, wait for it to dry and the flame dry it with a lighter...be careful, obviously isopropyl is flammable) and collect some spores. then just streak them across agar (once its settled in the petri dish, or whatever you use).
hope this helps.
-------------------- "Love says 'I am everything.' Wisdom says 'I am nothing.' Between the two, my life flows." Puget Sound Mycological Society
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Anonymous
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Re: multispore agar innoculation [Re: Blue Helix]
#2402425 - 03/04/04 11:30 AM (19 years, 10 months ago) |
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I don't have that kind of lab, or a hepa, and Isolation and testing works just fine. I am searching for the best isolates for my particular environmnet, throughout different times of the year. My home temps fluctuate during the seasons by about 10 degrees. Those isolates that perform best during the winter, perform not as well during the summer. Multispore results in average casings, which is better then poor, but not as good as above average. 10 original multispore plates can yield 50+ vigourous substrains to test. Of those fifty+ you might find 1-2 that are superior to the middle of the road. Instead of yielding 11 grams bone dry, you might be yielding 15 grams bone dry. an extra 4 grams per cup, is an extra ounce per 7 cups substrate. Now you can process far less substrate to get the same yields. Personally I don't like cleaning jars. The extra effort at the beggining, results in maintaining your Strains as close as possible to there oringinal state you obtained them in. It also enables you to have clones of known vigor, at known parameters, yielding greater consistency. Instead of having 1 average GT, you now have multiple GT clones that are above average, and varied from each other. You also still have the original print in storage, and prints from each generation you grow out from each clone. Hard to fruit species are hard to fruit becasue they require very specific conditions to fruit. Isolation won't increase the range of those conditions, but it can help you find those individuals that are skirting the edge of the range. IE, if the species fruits in a raqnge from 60-70 F, you might find the better producers at the 70F side of the range, which makes fruiting easier if you have trouble maintaing 65F temps, but can maintain 70F. This is SELECTION from a populaiton. This is how all breeding programs start, once pedigree is determined. Identifying the best individuals from the population for your specific requirements. If you are just happy getting fruits every time, AVERAGE is just fine. EDIT. I too am REALLY sensitive to shrooms. I use them very infrequently also. I don't feel superior to anyone or anything. I just like to see Facts remain facts, and not get obscured. The vested intrest you are refering to is TO STAY IN BUSINESS. To offer products that do not live up to CLAIMS will not keep you in business forever. I can assume you are refering to STAMETES and Fungi perfecti. The guy is still in business, and still sells CULTURES. He is offering cultures of KNOWN PRODUCERS. When someone is going to invest money in commercial growing of edibles/medicinals, they usually want to be able to take the Mushroom for granted. Ie. I am using this substrate in this environmnet, so I buy a culture that has been tested, and selected for inTHOSE EXACT ENVIRONMNETAL PARAMETERS. Then you just have to worry about maintaining those variables. I find it funny that you question the legitamicy of a practice that has been used since man began Agriculture. We selected for desirable traits, from large poluations of individuals. We did not select for PAR. In a natural poulation of Marijuana, the majority of individuals will be hermaphroditic, and average potency. Is this desirable if you like to smoke pot.
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Anonymous
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Re: multispore agar innoculation [Re: Blue Helix]
#2402496 - 03/04/04 11:57 AM (19 years, 10 months ago) |
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On a side note if you are interested in STRAIN VARIABILITY. Search a thread on Copelandia cystidia variation. It has alot of pictures that might interest you, as well as measurements. That thread is showing you visually just how variable a single species can be throughout the world.
That is morphological variation of a just pleurocystidia and spores only!!!
In the plant pathogenic fungal world, you can have a single species that has variants that only eat one type of tree, while other variants only eat another type of tree. One species, becoming multiple species becasue of host specificity. Just an Example of variability.
Having clones that colonize at 75 F, and others at 85 F. Ones that fruit at 85F and others that fruit at 65F. Doing so without any difference in time frames. Variability.
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daywalker
Do you dream in color?

Registered: 10/21/03
Posts: 783
Loc: in my skin
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Re: multispore agar innoculation [Re: Psilygirl]
#2404078 - 03/04/04 07:19 PM (19 years, 10 months ago) |
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works for me thanks for the tips!
-------------------- "I suggest we ... learn to love our ... selves before it ... becomes illegal."
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azurescens
member
Registered: 02/17/04
Posts: 717
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Re: multispore agar innoculation [Re: daywalker]
#2405043 - 03/04/04 11:26 PM (19 years, 10 months ago) |
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@cro
Is your good friend Billy Baldwin?? Because that is who that quote is from in the movie 'threesome'
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@cro
new name


Registered: 12/07/02
Posts: 1,224
Loc: The PNW
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Re: multispore agar innoculation [Re: azurescens]
#2405259 - 03/05/04 12:15 AM (19 years, 10 months ago) |
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I'm not sure what you're referring to, I didn't quote anything in my posts. And I definately don't know any Baldwins I try my best to avoid their movies. But as for sterilizing your utensil I would flame first then wipe with alcohol to cool it down and not kill the spores. Peace - @cro
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Scourge
The Dr.
Registered: 12/03/00
Posts: 293
Loc: Near flint Michgan, USA
Last seen: 18 years, 1 month
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Re: multispore agar innoculation [Re: ragadinks]
#2407025 - 03/08/04 12:38 PM (19 years, 10 months ago) |
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while not as good as a glove box IMO, adding h2o2 is a fairly safe method. there is little risk of contamination because h2o2, in proper concentration, will kill any bacteria and mold spores. the only real risk of contamination using proper moisture and h2o2 concentration would be if living mold got onto your substrate. not likely.
-------------------- "Those who walk in Love and Truth shall grow in Honor and Strength."
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@cro
new name


Registered: 12/07/02
Posts: 1,224
Loc: The PNW
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Re: multispore agar innoculation [Re: Scourge]
#2407101 - 03/08/04 12:52 PM (19 years, 10 months ago) |
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Yeah, peroxide's good at killing all types of spores, including cube spores. If it is a multispore inocculation peroxidated agar won't let anything grow. Although, if in the slightest of concentrations it could work and has I believe Ganja/Paid/Loki tried this and it worked for him.
Pea ce - @cro
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RogerRabbit
Bans for Pleasure


Registered: 03/26/03
Posts: 42,214
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Re: multispore agar innoculation [Re: mycohomme]
#2411739 - 03/09/04 05:35 PM (19 years, 10 months ago) |
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Another good trick is to cool the inoculating loop in the agar of the receiving dish. This will leave a trace amount of agar on the loop and help when you use it to collect spores. Just touch it lightly to the print and thousands of spores will stick to it. Personally, I try to bury the spores in the petri dish by rotating the loop into the agar in one spot in the very center of the dish. I disagree with spreading or streaking the spores around the dish. Try to keep them all in the center, so after germination they can do that 'mushroom sex' thing easily, then sector into the various strains you would isolate.
Perhaps what blue helix is saying is he's never seen a 'multispore' created project refuse to fruit unless contaminated. I have, but that's not the issue. I always isolate until there are no more sectors, then fruit(or try to) each substrain. By keeping masters, I can determine the best fruiting isolate, then go back to the refrigerator and only grow out that one for future cycles. I've done this with every strain in my library. If I isolate ten substrains(sectors) from a petri dish, one might fruit very well, two might fruit so-so the way multispore would, and it is very common that the other seven won't ever set a pin.
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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RogerRabbit
Bans for Pleasure


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 11 months, 2 days
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Re: multispore agar innoculation [Re: RogerRabbit]
#2411747 - 03/09/04 05:39 PM (19 years, 10 months ago) |
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Also, When using a spore syringe with agar, simply use one drop in the very center of the dish. Try to keep the dish flat when you wrap the parafilm around it to keep that drop right in the center. It will work better for you that way.
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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