  
Jesse James (Spacecowboy)
| | Posted on Friday, July 18, 2003 - 11:01 pm: | 
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This is the original Tek I got from Proff. PF; to help me with some experiments. My ultimate plan is to use it for the cultivation of wild morels that grow in my area. The Professor gave me permission to reprint it here at Mycotopia. I believe it may be a small except from the rumored “PF Tek 2,” if these is such a thing.
I highly encourage all of you to purchase the Professor’s new book on Stonehenge, METASPHERE - ARCHAEOSOPHY OF STONEHENGE, it makes complete sense, and gives new insight into “The meaning of life,” and why there is something and nothing. It will help to open your third eye. I read it, somewhat, and it is defiantly worth it.
It is hard to find words to explain the feeling you get once it sinks in. I know WHAT we are, and now I have a better understanding of WHY because it is written in the stones. At first I thought I had been brainwashed, but then I started talking about it with other people, from creationists to evolutionists, and they all seemed to agree. I feel as if I have had a revelation/epiphany. The entire book itself is very enlightening and it does a spectacular job of stating the archeological facts of Stonehenge.
For more info check out:
http://fanaticus.com/stonehenge.htm
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Roc (Rochester)
| | Posted on Saturday, July 19, 2003 - 01:16 pm: |
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Thanks for sharing...
Roc |
Hippie3 (Admin)
| | Posted on Saturday, July 19, 2003 - 02:00 pm: |
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ditto...
had not seen this before.
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Fanaticus (Fanaticus)
| | Posted on Saturday, July 19, 2003 - 09:36 pm: |
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There is a cut off of the text and some coma and quote sign snafues (using a lame mac) Here is a rewrite with corrected stuff.
PF PEROXIDE AND BROWN RICE CLONING TEK
by Professor Fanaticus
The original idea comes from Rush Wayne's cultivation manual "GROWING MUSHROOMS WITH PEROXIDE". The following is a simplification of the idea down to the smallest degree, using brown rice powder, peroxidated water dilutions and small jars.
MATERIALS
1. Small jars with lids
2. Brown rice powder
3. Regular store bought Hydrogen Peroxide (3%) antiseptic solution (usually in a brown bottle)
4. Dissection knife and long needle (exacto etc)
5. Living mushroom or fungus, or culture of fungus
As a preliminary, start reasonable clean, but the following is to be done in the open air, without sterile implements or containers.
Mix the peroxide antiseptic and water at a 20% ratio. Example - 2cc peroxide and 8cc water. Place an amount of brown rice powder into the jar and add some of the peroxidated water to get a slurry, or very wet condition. The diluted peroxidated water sterilizes the medium and the jar. For extra experiments, you should try more potent peroxide contents in the brown rice peroxide slurry medium (30% - 40% - 50%). It is so easy, it can't hurt to experiment and you can benefit from it.
Tear the shroom apart and with an exacto knife, excise a small fragment about the size of a match head. With a long needle, knock or scrape the shroom fragment off the exacto blade and place the fragment on the surface of the peroxidated brown rice slurry. If you are working with previously made plate or slurry cultures that have mold growing along with the pure white good fungi, follow this tek using a small fragment of the desired fungi excised out of the culture. If the culture grows back clean, that doesn't mean that it really is clean, because there will be contaminant spores in the mycelium (which is there anyway because of the open nature of this tek). When the culture is to be used PF style, when it is reslurried with fresh peroxide water, the contaminant mold spores and bacteria endospores will be killed while the growing fungi mycelium will survive to grow clean - PF style or any style you like.
Screw the lid tightly onto the peroxide slurry jar and put the jar in a warm place for growth. The culture can be opened and exposed for observation or experimentation without danger to the growing medium and culture.
Under magnification (10x), the fragment of mycelium appears to "melt". It also turns blue, as if it were killed. Within a few days, tiny white hair like tendrils (hyphae) will appear on the "melted blue" fungus. It will grow and fill the the culture jar.
To use the culture for further inoculations, add 20% peroxidated water to the culture, mix the brown rice and fungus, and with a syringe, draw up water and inoculate PF jars, with the standard PF spore syringe technique. This is done nonsterily also because the peroxidated water will kill contaminant spores. But flame sterilize the needle before injecting into the sterile PF substrate. Instead of flaming the needle, an even better way to sterilize the needle (outside of the needle) is to wipe the needle with a tissue soaked with rubbing alcohol. Let the needle briefly dry of the alcohol before injecting. Use the culture well before it grows in. That way, it is much easier to get a usable slurry for the syringe without clogging.
As an addition, try more potent peroxidated water ratios. For instance, a 50% ratio works also. That would be half peroxide antiseptic and half water. Increase peroxide content until the mycelium doesn't survive. Then back off the amount of peroxide and use a near death peroxide load for guaranteed clean results. But a 20% peroxide to water ratio seems to be perfect.
If you are working with previously made plate or slurry cultures that have mold or bacteria slime growing along with the pure white good fungi, follow this tek using a small fragment of the desired fungi excised out of the culture. Make sure you don't have any growing mold or bacteria in the good mycelium because the peroxide won't kill it (as it doesn't kill the good fungi). If the culture grows back clean, that doesn't mean that it really is clean, because there will be contaminant spores in the mycelium (which is there anyway because of the open nature of this tek). When the culture is to be used PF style, when it is reslurried with fresh peroxide water, the contaminant mold spores and bacteria endospores will be killed while the growing fungi mycelium will survive to grow clean - PF style or any style you like.
TEK PROBLEMS
The only problem is syringe needle clogging. As a remedy, do not allow the culture to fully grow in or get thick. Keeping the culture "thin", allows a good breakdown of the mycelial fragments for use in a syringe. At first, doing the tek can be messy, but learning is quick and easy. Finding and using needles as big as possible is important.
But another route is very possible and actually preferable. Using long glass bulb pipettes are very good to use, but here, one must customize and "tweak" the teks a bit (one must be careful when injecting PF style jars with the bigger pipetts and not to breach the top vermiculite contaminant barrier). Also, flaming the glass pipetts can sometimes break the glass. In this case, always sterilize the outside of the pipette with the rubbing alcohol. Bulb pipettes are actually better to use than needles and syringes because they don't have much problem with clogging as compared to needles and syringes with mycelial slurries. But if the Professor's tek is followed closely and the cultures are not allowed to grow in and get thick, the 18 gauge needles with 10cc syringes work OK.
PRINCIPLES OF THE PEROXIDE TEKS
Hydrogen Peroxide is a powerful antiseptic. The solution of Hydrogen Peroxide bought in a drug store is 3% Hydrogen Peroxide and 97% water. Even at this low concentration, and with further dilutions, the germ killing is potent. But that germ killing power only works for micro fungi spores and bacteria endospores. A micro-organism that has germinated into its secondary form (mycelium), is safe from the antiseptic power of diluted Hydrogen Peroxide. But the ungerminated spores, bacteria endospores, and microbes are all susceptible. If there is bacteria that is growing (germinated), it will not succumb to the peroxidated water (just as the fungi mycelium is not succumbing). Also, any mold that is growing will survive. A clean fragment of shroom flesh or mycelium from a mold contaminated culture has germs all over it, but only in the spore or endospore form. They won't germinate in the peroxidated medium and water or on the recovering mycelia.
The tek is like a tightrope act. The mycelium that is cultured in the peroxide enriched medium can survive and grow, but it is not clean. Any spores or bacteria that are "piggy-backing" on the mycelium and not in contact with the peroxidated medium can come to life if given the opportunity.
After the culture of mycelium grows, it can be rehydrated with more peroxidated water. The spores and bacteria endospores that are "piggy-backing" on the mycelium will die. The mycelium in its fully secondary form, will survive the new peroxidated solution, "cleaned".
BY PROFESSOR FANATICUS |
Hippie3 (Admin)
| | Posted on Saturday, July 19, 2003 - 09:40 pm: |
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thx
archive material |
John Doe (Smellmyfinger)
| | Posted on Saturday, July 19, 2003 - 11:02 pm: |
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So you Can use strain up spores with peroxide |
Joe Millionare (Rotterdam_Y2k)
| | Posted on Sunday, July 20, 2003 - 03:33 am: |
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My girlfriend speaks very little english. She would like me to ask whether she can see a picture of the "slurry?" Is there anyway someone can provide a recipe? 8cc water, 2cc h2o2, ?? brown rice flour. For those of us that use english as a second language, the term "slurry" is a bit ubiquitous...
--She wants me to let you know that she thinks this method is brillant.--
Dank u Mycotopia! Uw informatie is onschatbaar!
P.S. The more mushrooms she grows, the more whoopie I get to make!
Thanks! |
Fanaticus (Fanaticus)
| | Posted on Sunday, July 20, 2003 - 04:09 am: |
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By John Doe (Smellmyfinger) on Saturday, July 19, 2003 - 11:02 pm: Edit
So you Can use strain up spores with peroxide
answer from the Professor --- no - only mycelium that you get from a shroom or a culture or a cake or whatever. You know, that white fluffy stuff.
By Joe Millionare (Rotterdam_Y2k) on Sunday, July 20, 2003 - 03:33 am: Edit
My girlfriend speaks very little english. She would like me to ask whether she can see a picture of the "slurry?" Is there anyway someone can provide a recipe? 8cc water, 2cc h2o2, ?? brown rice flour. For those of us that use english as a second language, the term "slurry" is a bit ubiquitous...
--She wants me to let you know that she thinks this method is brillant.--
Answer from the Professor - a slurry made from brown rice powder for the culturing looks almost like an agar culture except that it is not "hard". It is a messy kind of thing. The recipe is wide open for variation. When you do your final slurry for injection, then it is a brownish liquid with mycelium and brown rice powder floating around in it - again, the recipe and amounts of water to be used vary greatly. You just need to try it. With a fresh shroom or culture, you can do zillions of them to see how you like to do it. There are no definitive rules on this one.
But guess what? This tek can elliminate having to buy any spore syringes. But it lasts only as long as the culture will remain unsenescent, and I have heard all kinds of claims about this.
But then, if you take a spore print and you make a culture with it and it is contaminated, you can use the tek anyway. But you need to have some clean mycelium growing amongst the contaminants. If you got bacteria, this tek won't help. Then you should try that antibiotic agar agar (which can't stop mold - only bacteria) you can get from Fungi Perfecti, and if you have mold, then you can use this tek to isolate and get your culture clean and ready for PF style.
Professor PF |
Joe Millionare (Rotterdam_Y2k)
| | Posted on Sunday, July 20, 2003 - 04:53 am: |
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My girlfriend suggests that the unsenescent culture could be transferred via syringe (14 guage needles work best) to the KARO Tek creating mass mycelia.
Dank u zeer, Professor! |
John Doe (Smellmyfinger)
| | Posted on Sunday, July 20, 2003 - 10:55 pm: |
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Well I know that you can use Spores on peroxided agar.... and thats a fact. |
nilla (Nilla)
| | Posted on Sunday, July 20, 2003 - 11:20 pm: |
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This is a shot in the dark but does anyone think its possible to do this idea with pf jars by adding this ratio of peroxide to the 1/4 cup of water per jar,this being tissue shot and skiping the jar boiling? |
jim dillows (Jimdillows)
| | Posted on Monday, July 21, 2003 - 02:14 am: |
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10:1 water:peroxide is a great ratio if you have solid tissue. For example, if you add water and peroxide in a blender to make a slurry for a syringe full of live tissue.
If you have to hydrate something like a casing layer or wood spawn 7 cups water: 2 tbs peroxide or 7/8 cup of water :1 teaspoon of peroxide.
(See Rush Wayne's 8 minute no pc spawn)
Peace
JD |
Hippie3 (Admin)
| | Posted on Monday, July 21, 2003 - 02:43 am: |
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peroxide kills spores, smellyfinger.
care to prove your claim ? |
PissyBee (Pissybee)
| | Posted on Monday, July 21, 2003 - 08:11 am: |
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Well, I'm no agar expert but you transfer a wedge of the cleanest cut of MYCELIUM to the peroxidated agar, not spores and as it says in PF's write-up, "A micro-organism that has germinated into its secondary form (mycelium), is safe from the antiseptic power of diluted Hydrogen Peroxide." That is the point of peroxidated agar and is the same principle that this method is using. Hope that helps.
PB |
John Doe (Smellmyfinger)
| | Posted on Tuesday, July 22, 2003 - 12:16 am: |
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Quote:Peroxidated agar is about the only place you can use spores and peroxide together. Of course this has been known for some time, but I see it needs reiterating.
A relatively heavy streak of spores will in fact germinate on peroxidated agar. I know, I didn't believe it myself, I think it was Major Millet back at the drool donkey who brought it to my attention. Even the peroximan himself, mr. wayne has been turned on to the idea by now.
I guess that a heavy streak of spores protects the spores in the middle from the peroxide, they germinate, and can immediately digest the peroxide. Of course, you should use the absolute lowest concentration of peroxide you can get away with.
By mycofile
(This is from Freinds, from another board and thought that we all should share as much info as possible. I have tryed it. It does work. Peroxide is getting to be the way to go.IMO... oh and nice to see you around pissy |
Hippie3 (Admin)
| | Posted on Tuesday, July 22, 2003 - 01:15 am: |
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interesting, somehow even after all this time i hadn't heard that. not too surprised millet never told me.
i'll do some digging now and see if i can further verify that.
know any urls of threads on boards that cover this ?
the quicker i see a few others confirm the statement the quicker i can apologize for contradicting you and begin correcting my advice to reflect your input.
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John Doe (Smellmyfinger)
| | Posted on Tuesday, July 22, 2003 - 07:18 am: |
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Hope This help...Im shure its not in a text book at the library yet 
http://www.nansnook.com/forum/index.php?act=ST&f=21&t=9236&s=697752a5bf31041f84d7cbe9311c0364 |
Hippie3 (Admin)
| | Posted on Tuesday, July 22, 2003 - 12:27 pm: |
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ok, i failed to realize you were already "quoting" mycofile.
i can see how it would work.
at first, there would be a violent [microscopic] reaction between the spores and the peroxide.
they would mutually obliterate each other.
that's how peroxide works, in killing the spores it destroys itself, like a matter-antimatter reaction.
so the trick would be to concentrate enough spores in an area to overwhelm the peroxide and survive with enough spores left over to germinate in the now peroxide-free zone.
once germinated of course the remaining peroxide would not be a problem.
of course, the same principle would apply to any contams present, so a heavily contaminated print done in this manner could still grow out contaminated.
thx for bringing this to my attention,
from the responses to that thread i can see that i'm not the only one who was unaware of this phenomenon.
i apologize for contradicting you before i knew all the facts, your help is appreciated. |
Hippie3 (Admin)
| | Posted on Tuesday, July 22, 2003 - 12:36 pm: |
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let's toss in fungusmaximus' version of mycofile's peroxidated agar formula, it's in the archives too but it helps if the info's in multiple places.
Quote:The formula I use for H202 agar is one the mycofile posted at mycotopia a long time ago. Base formula for smaller batches of agar.
3 grams agar
3 grams dextrose
149mls h20 potato water etc.
add 1ml h202 @ 130*
The max amount that mycelium will grow on without being severely
stalled is 7ml h202
You must compensate the addition of h202 by subtracting from the amount of h20.
note: the h2o2 [peroxide] is NOT added until after sterilization, as the agar begins to cool down [when it reaches 130°F.]
archive material |
not *really* a Jedi (Mycofile)
| | Posted on Wednesday, July 23, 2003 - 11:14 pm: |
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I meant to update that thread from the shroomery. Wayne mentions using spores on peroxidated agar in his peroxy manual volume II. He seems hesitant to recomend it, even stating a vague fear about genetic damage at this stage. But he does say that it works, just take care to make a heavier "streak" than normal. In my and others experience, it's a little more hit or miss than using mycelium, but certainly no genetic damage is evident. Regardless, Wayne is considered the peroxide guru, so his statements should be backing enough for the claim.
No offense against anyone, we are all guilty of it, but this is a perfect example of people repeating what they've been told without trying it for themselves. Everybody thinks peroxide kills spores, which it does in theory, but not practice. At least not in all circumstances. I would still hesitate to use spores on a peroxidated liquid, (but again, I haven't tried it, so I might be leading the witnesses), and I think it would work better on the firm suface of agar than on a BR slurry or paste.
On the original post, I'm surprised more people aren't familiar with this cloning tek. It was popularized by several posters a few years ago calling it "brown rice paste", and the professors version was on his website for sometime before his unfortunate bust. |
Hippie3 (Admin)
| | Posted on Wednesday, July 23, 2003 - 11:31 pm: |
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it seems to me that actually peroxide does kill spores 'in practice',
but like all other forms of sterilization,
it has it's limits.
if you overwhelm it, which is essentially what this does, it's still killing spores, just not every single one. it may kill 900,000,000 spores and leave only 100 alive but 100 left is more than enough to get the show underway.
i do remember the paste method, i just didn't associate it with PF, my memory must be slipping.
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not *really* a Jedi (Mycofile)
| | Posted on Wednesday, July 23, 2003 - 11:51 pm: |
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Sure, but what I meant was that the common notion that you can't use sprores with peroxide is false. Everybody (including myself) repeated what we were told based on someone elses statement of their theory, not what we knew from experimental evidence. |
Hippie3 (Admin)
| | Posted on Thursday, July 24, 2003 - 12:04 am: |
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well i wouldn't feel too badly about going by what we were told instead of re-inventing every wheel, that would slow us down alot. blame instead the ones who told us.
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Yachaj Payé (Yachajpb)
| | Posted on Friday, July 25, 2003 - 10:59 am: |
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Spore germination on peroxidated substrates is ineed possible but also risky (may cause genetic damage). A more useful method is to germinate the spores in a test tube with a solid substrate (a slant). In an unsterile environment the opening of the tube just needs to be flamed.
As long as the mouth of the tube is hot enough contaminants will not enter. You can add spores or a mature piece of mushroom gill by using an inoculation needle (or metal wire).
This method is very old, popularized by Bob Harris in the nineteen seventies. It is also in Waynes' 2nd book.
Yachaj |
Hippie3 (Admin)
| | Posted on Sunday, July 27, 2003 - 01:19 pm: |
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thx, yachaj.
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Imok Urok2 (Imok)
| | Posted on Monday, July 28, 2003 - 12:00 am: |
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I think PF talked about using that tek himself.
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Alien Primate (Alienprimate)
| | Posted on Friday, August 22, 2003 - 09:16 pm: |
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So you could probably mix some of this rice flour/peroxide slurry (the extra runny stuff) and put it into a PF jar that was just birthed so that the mycelia still clinging to the jar will colonize the liquid, then you could suck it out after a week or two with syringes.
Cool deal, this will help open the door to cloning for some guy I know.
Thanx so much PF! |
Hippie3 (Admin)
| | Posted on Saturday, August 23, 2003 - 03:30 pm: |
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do that inside a glovebox [the birthing, too] and it'd usually work out. |
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