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dupee419
Farmer

Registered: 05/09/04
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Hybrids
#2656522 - 05/09/04 11:40 AM (19 years, 8 months ago) |
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I was pondering how to make a hybrid strain of shroom and I can't find any information on it at all. Does anyone here know anything on the Manual hybridization of Psilocybin containing mushrooms? I'm tempted to try my hand at it if I can find enough information.
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ATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
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Re: Hybrids [Re: dupee419]
#2657989 - 05/09/04 06:58 PM (19 years, 8 months ago) |
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The FAQ...
Good Luck!
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Mycena
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Re: Hybrids [Re: dupee419]
#2659188 - 05/10/04 10:21 AM (19 years, 8 months ago) |
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highly technical
you can try and isolate monokaryons - youll need a micrscope t look for clamp connections. you label them then try crosses
ths is good within species
between species your looking at protoplasts cells are treated with enzymes to remove the wall and kept in a buffer solution, then they can be fused using a few techniques and a small amount can regenerate as dikaryotic interspeccific or intergeneric hybrids most will be freaks but some work - like a cross betwee Shiitake and The golden oyster that made golden shiitake Further selction can then proceed Intrageneric crosses might be better
take for example an Azurescens/cyanescens hybrid, or a Cubensis/ Stropharia rugosoannulata hybrid (perrennial cubes?)
theres a lot of potential if you can pull it off
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Transplant
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Re: Hybrids [Re: dupee419]
#2664724 - 05/11/04 06:23 PM (19 years, 8 months ago) |
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Fuse them.
"Mycelium starts to grow when mushroom spores germinate. Unlike seeds, which have diploid cells andfully formed embryos, mushroom spores are haploid cells, either + or - . You can?t tell one strain from theother and either type will grow into mycelium. However, in order for mushrooms to form, + and ? mycelium must grow together and fuse. This happens in nature largely by chance, but mushroom growerscan force the issue by growing spores of the two strains together in the same culture dish. Once the fusion occurs, the ?new? culture can be maintained by way of tissue culture. Typically, a tiny piece of tissue isremoved from the upper stalk of a mushroom where the stalk meets the gills. The tissue fragment isplaced on a nutrient agar medium in a petri dish. The same agar media is used that would be used to growmycelium from spores. Within a few days to two weeks the mycelium will grow and cover the agar in a petri dish"
http://www.cce.cornell.edu/clinton/ag/forestry/mushrooms/Growing%20Gourmet%20Mushrooms%20bw.pdf
-------------------- Will Screw For Shrooms
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doc34
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So basically what you are saying is that you can innoculate a jar with two different strains(pesa-gt)and "if"they mate +pesa and -gt then you would have a hybrid(I know,what are the chances in that happening)?I'm just speaking theoretically here ok? Then you can clone the hybrid to maintain a new strain?
I probably sound stupid,but its just a question.
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Mycena
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Re: Hybrids [Re: doc34]
#2695953 - 05/18/04 12:36 PM (19 years, 8 months ago) |
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Chances of success are so low as to be not viable and certainly not distinguishable
you should take two very different cubes like say Penis envy and anything else isolate monokaryons on agar then test a number of matings
maybe you will get something, at the least you might put some life back into PE
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Anonymous
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Re: Hybrids [Re: doc34] 1
#2697915 - 05/18/04 05:38 PM (19 years, 8 months ago) |
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In theory you could inoculate a single jar with spores of two different strains and potentially grow out a third new strain.
The problem with this approach is it is not controlled. Spores tend to clump and mate with the other spores in closest proximity, so two spores from different strains may never come into contact. Also you are multisporing, so MANY substrains are being formed, and differentiating between the dikaryons formed will be next to impossible, when they are all growing together in a mixed tray. You might see a difference, but how can you prove it without doing some kind of DNA anylisis, or by doing compatability studies.
Isolating monokaryons from a verry dilute germination, and doing controlled matings results in more verifiable and reproducible results.
First generation spores from hybrid crosses can contain many lethal alleles, and germination rates can be very low. After a single generation the rates rapidly increase.
If a hybrid was formed, then 3/4 of the monokaryons from the new strain will mate with either of the two donor strains, becasue they will share a common ancestor. It is a new strain becasue you have created spores that contain combinations of genes that did not exist together, before.
If I start with GT and EQ, and they are not related at all. I create a hybrid Strain GQ . this GQ will share a common mating type loci with both EQ and GT. So The GQ will mate with the EQ at 75%, and the GQ will mate with the GT at 75%. The EQ will mate with the GT at 100%.
All of these matings will be done on agar, and tested for fruiting by conventional means. Then you have to select for the best individuals, and clone those for use. Or maintain the responsible monokaryons for the superior indivdual dikaryons, and mate them every time you want to produce the KNOWN exceptional dikaryon expressing the traits you want. If you start over with the spores produced from the second generation of the NEW STRAIn inbreed with itself, you are going to produce spores as variable as the spores produced by inbreeding either the GT or the EQ.
The name of the game is SELECTION, and Cloning for superior results. If you want to try and improve on what exists, you have to mate what is out there in new combinations and SELECT the best for future use(CLONES).
Any time you multispore, you have variability between the resulting dikaryons, but also similarity. Similar as in they all kinda resemble each other(shared traits) and variable in that they tend not to be identical to each other. One dikaryon may perform optimally at one temp, and another at another temp, etc... One may produce the highest average activity, while another may produce at the lower range. Similar in the sense that ALL the Dikaryons may produce slightly higher then normal Activity, the AVERAGE of the population of your hybrid Strain may be slightly higher then the Average of the two donor Strains.
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doc34
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Re: Hybrids [Re: ]
#2698054 - 05/18/04 06:13 PM (19 years, 8 months ago) |
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Thank you Masta'son,thank you!
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Sterile
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Re: Hybrids [Re: ]
#2700036 - 05/19/04 05:41 AM (19 years, 8 months ago) |
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So do we actually have any good results that you know about? Are there any good hybrid strains out there?
Did anyone try to mate Cubes with a more potent shroom and actually have results?
-------------------- The Source Of The Force Is The Power Of The Mind "if you don't like what you're doing, you can always pick up your needle and move to another groove." - timothy leary" Anno: "-I can do anything with those clouds!" Annos Tek
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Anonymous
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Re: Hybrids [Re: Sterile]
#2705068 - 05/20/04 10:12 AM (19 years, 8 months ago) |
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Did you read the posts.
There have been no scientific studies done on compatability between strains of cubensis.
You cannot by conventional breeding systems, mate one species with another species. So no one has tried to mate a cubensis with a more potent species.
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RogerRabbit
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Re: Hybrids [Re: ] 1
#2718497 - 05/23/04 01:31 PM (19 years, 8 months ago) |
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As an alternative to putting two types of spores in a jar, try inoculating two different isolated strains of one species onto the same agar wedge. Simply transfer two wedges of myc to the same plate. When the two grow together, you'll see a 'line' form between strains. Sometimes one strain will gobble up and 'eat' the other one. Sometimes, each holds its own turf and the line stays put between them, where each has its own territory on the petri dish. About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain. It is definately and absolutely possible, and I've done it. Next fall, when I have time to resume the hobby, I'll do it again and write a photo documented tek. Until then, anyone who has time and inclination to experiment, now knows how to go about it.
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JohnHolliday
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Hybridization is not only possible, it is common and done every day in the commercial mushroom industry. The white button mushroom you see in the store are hybrids. The easist method is somatic fusion with a hybridizing agent, such as Snake venom as detailed in the article at http://alohamedicinals.com/cordy_IJMM_hybrid_article.pdf The coprophytic mushrooms such as cubensis are the easiest of all to grow and they readily hybridize. There are a number of techniques, such as spore series dilution, followed by liquid culture (preferably, although solid substrate can be used) and grow out to see what the strain characteristics are like. That is quite time consuming though. Perhaps it is more practical to just adjust the substrate/culture parameters to modify the analytical signature of the final product. Read the article on rye grass chafe as an amendment at http://www.alohamedicinals.com/sdarticle.pdf If you follow this technique, you will find that you have increased your desired results perhaps 10x to 20x.
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Jeremy_Davis
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Registered: 04/22/05
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Hey John! I'm soooooo excited to see you over here! I know this is gonna be interesting having you around! Light and Love, Jeremy Davis
-------------------- Jeremy Davis Educational Concerns for Hunger Organization, Inc. Check out the ECHO mushroom blog page to see our lab, growing facility, and more-www.echotech.org/greta
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whiterasta
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Registered: 04/09/02
Posts: 1,780
Loc: Oregon
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Quote:
There are a number of techniques, such as spore series dilution, followed by liquid culture (preferably, although solid substrate can be used) and grow out to see what the strain characteristics are like. That is quite time consuming though. Perhaps it is more practical to just adjust the substrate/culture parameters to modify the analytical signature of the final product.
spore series dilution and liquid microculture can give genuine intraspecies hybrids such as the GQ mentioned above.It would take hundreds of microcultures being fruited to determine the rate of fusion between variety and to gain a stable sporulating hybrid. Interspecies such as cube and cyan are not compatable under normal conditions but cell biology has progressed to the point a sterile mule could be produced relatively easily in a lab. But what would it's substrate be? Were I to contemplate a cross it would be an azurescens with a warmer weather, more cosmopolitan wood lover. Any suggestions on the warm wood lover? WR
-------------------- To old for this place
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RogerRabbit
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I think it's jumping the gun to assume that a fungal cross-species hybrid would be sterile because that's what happens with mammals. Work is underway to test this theory. RR
-------------------- 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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whiterasta
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I was referring to what could be readily acomplished now, as you say testing for interspecies fertility is ongoing however the substaces used to induce genetic intermingling, which I would assume to be the available method,are not conducive to an orderly blending of genes and may not produce something fertile. I was not making my assumption on mammilian genetics. Any suggestions on a warm weather wood lover?  WR
-------------------- To old for this place
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RogerRabbit
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Reishi. RR
-------------------- 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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whiterasta
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So in the topic of this post how does reishi relate? I apologize if I have misunderstood your answer but how in god's name does Ganoderma equate to a warm weather wood loving Ps. species? If you are just being a smart ass then...HAHAHA WR
-------------------- To old for this place
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WhiteBunny
How deep doesthe rabbit hole go?


Registered: 07/29/05
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Roger do have you continued to grow this hybrid that you made? Did you fruit it and if so what were the results?
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whiterasta
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Registered: 04/09/02
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Loc: Oregon
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Quote:
About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain.
LOL I suppose it could not be just intermingled compatable mycelia as dikaryotic pairings are impossible. Only monokaryons can 'pair"and if you have already paired them and are using agar wedges of dikaryotic mycelia, tell me how this "hybrid" occurred? RR you are usually quite knowlegable but this time you are full of Merde`. I am sure the holes in your reasoning are quite apparent to most on this issue.Spore dilution and hundreds of microcultures are how fungal hybrids are produced not growing dikaryotic mycelia in proximity. WR
-------------------- To old for this place
Edited by whiterasta (10/08/05 11:54 AM)
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RogerRabbit
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Quote:
whiterasta said:
Quote:
About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain.
LOL I suppose it could not be just intermingled compatable mycelia as dikaryotic pairings are impossible. Only monokaryons can 'pair"and if you have already paired them and are using agar wedges of dikaryotic mycelia, tell me how this "hybrid" occurred?
Sorry my friend, but it is you who are mistaken. Dikaryons are capable of hybridizing. In fact, I learned the exact procedure I mentioned above from paul stamets at his master's seminar. I have no idea the exact process beyond the mechanics of setting up the experiment because it's the mycelium that does all the work. The grower simply places the two sets of dikaryons on the same petri dish and then sits back and watches. One must be very careful at this stage of human understanding of how fungi work when using phrases such as "only monokaryons can pair", because such reasoning is incorrect. RR
-------------------- 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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EonTan
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Registered: 08/18/04
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ANASTAMOSIS Dikaryon to Dikaryon mating. Bisporous mushroom spores have both nuceli present and hybrids formed are via this route. IE Agaricus bisporous.
Conventional breeding programs with heterothallic mushrooms tend to be more controlled via MONOKARYON to MONOKARYON mating. The frequency of mating will be VERY HIGH the more unrelated the strains within a species. Lethal combinations may need to be worked out in the first generation of spores produced. So your actual germination/mating rates may be low.
Species to species and Genus to Genus, would require techniques that bypass Compatablity systems. IE protoplast isolation and fusion.
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youngbae
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Re: Hybrids [Re: EonTan]
#4835675 - 10/21/05 06:57 PM (18 years, 3 months ago) |
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i'm trying to make new hybrid...but...it's very difficult... since i've been trying to nearly 1 year.... so little result i obtained.... but the compensatory will enough....
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JJ01769
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Re: Hybrids [Re: youngbae]
#4848979 - 10/25/05 02:33 AM (18 years, 3 months ago) |
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I dont understand this stuff exactly. Im still learning all the LC and agar things but I was thinking. What about using two different LC from like the Karo/water and put them in a WBS jar? This maybe completely off and everything but it just popped in my head. Possible to work?
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RogerRabbit
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Re: Hybrids [Re: JJ01769]
#4849297 - 10/25/05 05:55 AM (18 years, 3 months ago) |
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You would have fruits from two different strains. You could even do it with different species. They will very rarely hybridize that way. RR
-------------------- 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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Zen Peddler


Registered: 06/18/01
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why wouldnt strains of cubensis be compatible? After all the delineation of a species often relates to its mating compatibility. At the end of the day a hydrid of two strains of cubensis is still the same species - so its not a hybrid, just a new substrain of that species. The likelyhood of this being successful between different SPECIES as opposed to strain is FAR more challenging. If you think it will be done in your back room I think your probably dreaming.
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RogerRabbit
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Correct, strains of the same species are much more compatable and dikaryotic/dikaryotic pairings are also possible, but not likely. It will happen occasionally however. I've also put different species in the same substrate before. For example, you can mix copelandia and cubensis in the same manure or straw tray and they'll fruit at the same time. I've never seen they hybridize this way however. RR
-------------------- 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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WhiteBunny
How deep doesthe rabbit hole go?


Registered: 07/29/05
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Quote:
RogerRabbit said: As an alternative to putting two types of spores in a jar, try inoculating two different isolated strains of one species onto the same agar wedge. Simply transfer two wedges of myc to the same plate. When the two grow together, you'll see a 'line' form between strains. Sometimes one strain will gobble up and 'eat' the other one. Sometimes, each holds its own turf and the line stays put between them, where each has its own territory on the petri dish. About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain. It is definately and absolutely possible, and I've done it. Next fall, when I have time to resume the hobby, I'll do it again and write a photo documented tek. Until then, anyone who has time and inclination to experiment, now knows how to go about it.
Where you able to fruit this hybrid and if so have a print or LC made?
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RogerRabbit
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Yes, and yes. Dikaryon to dikaryon pairings are quite common in the commercial mushroom industry. Let's say you have one strain of agaricus that has large meaty fruitbodies, and another strain that gives prolific flushes. Dikaryon-dikaryon pairings in this case would be called for in an attempt to get the best of both worlds. RR
-------------------- 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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WhiteBunny
How deep doesthe rabbit hole go?


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can I askj what were the two strain that you used to get the hybrid?
WB
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shoeareyou
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that would be absoloutely fantastic RogerRabbit Go on my son =)
Quote:
RogerRabbit said: As an alternative to putting two types of spores in a jar, try inoculating two different isolated strains of one species onto the same agar wedge. Simply transfer two wedges of myc to the same plate. When the two grow together, you'll see a 'line' form between strains. Sometimes one strain will gobble up and 'eat' the other one. Sometimes, each holds its own turf and the line stays put between them, where each has its own territory on the petri dish. About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain. It is definately and absolutely possible, and I've done it. Next fall, when I have time to resume the hobby, I'll do it again and write a photo documented tek. Until then, anyone who has time and inclination to experiment, now knows how to go about it.
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