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Lurksponge
Biochemist
Registered: 09/11/08
Posts: 44
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LC Cloning vs Isolating
#9370491 - 12/05/08 03:34 AM (15 years, 3 months ago) |
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I know the two are quite different, but in terms of what they accomplish (artificially selecting prolific fruiters/fast growers) are they just two paths to the same goal?
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J3illy
Trainee
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Re: LC Cloning vs Isolating [Re: Lurksponge]
#9370498 - 12/05/08 03:40 AM (15 years, 3 months ago) |
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Well from everything I've read, they're pretty similar, and both should achieve similar results. But cloning a fruit could still have multiple strains, whereas isolating is only 1 strain. I'm not sure how likely it is for a fruit to have multiple, but it's definitely possible as RR has pointed out.
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dead
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Re: LC Cloning vs Isolating [Re: J3illy]
#9370502 - 12/05/08 03:45 AM (15 years, 3 months ago) |
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Well, anyway when you clone a fruit you will know all the substrains in it will be able to produce fruits. There won't be any "impotent" strains...
-------------------- "The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows." Links: Nibin's Guide for Noobs some easy teks on bulk & grain prep. (my journal)
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J3illy
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Re: LC Cloning vs Isolating [Re: dead]
#9370511 - 12/05/08 03:50 AM (15 years, 3 months ago) |
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Yea from grows I've seen, cloning works out really well. You definitely know what you're gonna be getting, and w/ a nice even pin-set.
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Lurksponge
Biochemist
Registered: 09/11/08
Posts: 44
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Re: LC Cloning vs Isolating [Re: J3illy]
#9370531 - 12/05/08 04:07 AM (15 years, 3 months ago) |
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Thanks for the speedy responses! I was expecting as much, but just wanted to verify. Have an insanely rhizomorphic jar going right now and was going to case n fruit it, then take the biggest of the lot and clone that into some LC for further batches.
Once you've knocked up all your jars with the LC from the clone, can you just do g2g thereafter? Or would it be beneficial to measure the size of the fruit and continuously select larger fruits for further cloning?
-------------------- I need rising sound … And when it comes to that fantastic note where the rabbit bites its own head off, I want you to throw that fuckin' radio into the tub with me! My newb experiences and tips to other newbs
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ohmatic
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Re: LC Cloning vs Isolating [Re: dead]
#9370586 - 12/05/08 05:02 AM (15 years, 3 months ago) |
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Quote:
dead said: Well, anyway when you clone a fruit you will know all the substrains in it will be able to produce fruits. There won't be any "impotent" strains...
there are no "substrains in a fruit" - you isolate for this very purpose of CLONING the ONE AND ONLY genetic makeup of THAT ONE organism.
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
Edited by ohmatic (12/05/08 05:03 AM)
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veda_sticks
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9370606 - 12/05/08 05:26 AM (15 years, 3 months ago) |
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Quote:
ohmatic said:
Quote:
dead said: Well, anyway when you clone a fruit you will know all the substrains in it will be able to produce fruits. There won't be any "impotent" strains...
there are no "substrains in a fruit" - you isolate for this very purpose of CLONING the ONE AND ONLY genetic makeup of THAT ONE organism.
Isnt it still possible to have cloned tissue sector though??
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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ohmatic
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Re: LC Cloning vs Isolating [Re: veda_sticks]
#9370615 - 12/05/08 05:34 AM (15 years, 3 months ago) |
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Quote:
veda_sticks said:
Isnt it still possible to have cloned tissue sector though??
what ?
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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veda_sticks
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9370618 - 12/05/08 05:36 AM (15 years, 3 months ago) |
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Quote:
ohmatic said:
Quote:
veda_sticks said:
Isnt it still possible to have cloned tissue sector though??
what ?
Not that long ago someone said that a mushroom fruitbody growin from multispore could be made up from more than 1 strain of mycelium and that when cloned could produce sectoring though rare.
Cant remember who, but have heard a few people repeat this. Obviosuly the misinformation that i havent spotted then.
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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ohmatic
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Re: LC Cloning vs Isolating [Re: veda_sticks]
#9370620 - 12/05/08 05:38 AM (15 years, 3 months ago) |
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2 spores meet, produce dicaryotic myc and this eventually produced a fruitbody. its got one genetic makeup.
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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Nibin
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9370621 - 12/05/08 05:39 AM (15 years, 3 months ago) |
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Quote:
ohmatic said:
Quote:
dead said: Well, anyway when you clone a fruit you will know all the substrains in it will be able to produce fruits. There won't be any "impotent" strains...
there are no "substrains in a fruit" - you isolate for this very purpose of CLONING the ONE AND ONLY genetic makeup of THAT ONE organism.
A fruits are known to sometimes be made up of more than one strain. You still have to sectorize on agar to make sure you have a single strain isolate.
Also, another problem of cloning vs sectoring on agar is that your master tissue will already be over a month old if you clone, and that will limit the amount of transfers you will be able to make before senescense starts to appear.
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ohmatic
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Re: LC Cloning vs Isolating [Re: Nibin]
#9370644 - 12/05/08 05:55 AM (15 years, 3 months ago) |
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proof, i demand it !
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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HybridprX
Biodegrader of coir
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9370668 - 12/05/08 06:16 AM (15 years, 3 months ago) |
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I've never seen a tissue clone sector on agar myself...only one substrain need apply during fruit body production.
Awhile back when I first got into agar work I asked if I could take a clone from a prolific fruiting multispore inoculated grain/coir tub and then isolate it out even further on agar, the universal response was, no.
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Nibin
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9370831 - 12/05/08 07:45 AM (15 years, 3 months ago) |
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Quote:
As for cloning vs strain isolation, they're not related. By the time a substrate fruits, hundreds or perhaps thousands of strains have exchanged DNA, either weakening or strengthening the mass. What you get is a 'heinz 57' that may or may not be that great because the weaker genes and the stronger genes(mycelium) have all combined. An example would be mixed breed dogs. We've all seen good examples and others that are dumber then hell.
Strain isolation on agar begins when the spores first start to germinate. I make the first transfers as soon as I can see mycelium growing from the point of inoculation, long before sectoring can be detected. By doing this, and by continuing to separate each individual growth, you can isolate mycelium prior to the process of anastomosis combining dikaryons into a single mass.
You don't isolate looking for one super rhizomorphic strain. You isolate down to single sectors and then fruit out each one to determine the best performer. When you transfer mycelium to a grain master, the original petri dish the mycelium was taken from is placed into a clean refrigerator. By doing this, when you find the best performing strain, you then go back to your well marked petri dishes, thus your original P1 culture. This petri dish can be used to inoculate a few test tube slants that can be incubated for a week, then placed in cold storage. Whenever you need mycelium, a tiny piece the size of a grain of rice can be taken from the test tube and put on agar to grow out, while the test tube is placed back into the refrigerator. These stored test tube cultures preserve the low P value of your isolated strain for years.
I have a complete video tek on strain isolation and master slant preparation and use already filmed. I'll release it when I get the rest of the teks filmed, and editing completed. Hopefully soon. RR
Quote:
Yes. More than one strain can be present in an individual fruit. Thus, a clone is not an isolated strain, unless it was grown from an isolated line of mycelium. RR
It's not only me that says this.
-------------------- Newcomers guide-----> For all things shroomy
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ohmatic
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Re: LC Cloning vs Isolating [Re: Nibin]
#9371026 - 12/05/08 08:54 AM (15 years, 3 months ago) |
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interesting cheers.
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HybridprX
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Re: LC Cloning vs Isolating [Re: ohmatic]
#9371237 - 12/05/08 09:50 AM (15 years, 3 months ago) |
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amen, should have made the attempts then.
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Lurksponge
Biochemist
Registered: 09/11/08
Posts: 44
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Re: LC Cloning vs Isolating [Re: HybridprX]
#9372565 - 12/05/08 01:30 PM (15 years, 3 months ago) |
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Most excellent. Thanks again for the clarification. Looks like LC cloning only helps you get more consistant results from your initial starting culture whereas isolation weeds out the crap before it has a chance.
Makes me want to pickup some new syringes/spore prints and start on some agar work. It's interesting how when you start out this hobby it's all about just getting things to work. Then the obsession grows for perfection and further experimentation.
Thanks again for all the info guys. I appreciate all the help the shroomery has given me thus far in my hobby.
-------------------- I need rising sound … And when it comes to that fantastic note where the rabbit bites its own head off, I want you to throw that fuckin' radio into the tub with me! My newb experiences and tips to other newbs
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Nibin
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Re: LC Cloning vs Isolating [Re: Lurksponge]
#9372881 - 12/05/08 02:17 PM (15 years, 3 months ago) |
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In either case, cloning is way better than starting from multispore.
If you make a big LC from a good fruit you can inoculate pounds and pounds of substrate before you have to start off from spores again.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: Nibin]
#9373999 - 12/05/08 04:52 PM (15 years, 3 months ago) |
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This question is not intended derogative, so please don't take it as such, but is RR supposed to be some mushroom "God" or "Guru" or something? I mean, everyone quotes him and ONLY him when someone asks for proof. Is it just because he can intelligently state his opinions (and make them sound more proven), or is there literally no one more experienced/knowledgeable here?
Again, don't flame me on this--it's not meant to be insultive, just an honest question. I'm obviously not to "deep" into the mushroom community. ^_^
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J3illy
Trainee
Registered: 10/18/08
Posts: 3,344
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He's a veteran pro mushroom guru, and a Mod here. Any advice he gives is totally legit. He's probably grown, tested, tried, researched everything out there. He designed a totally automated FC - the Shotgun tub, and put out some DVD's titled "Lets Grow Mushrooms" which seem to be pretty popular.
If you wanna see him goto www.mushroomvideos.com
It's not to say no one else knows what they're talking about - but he's revered around here, lol.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
#9374171 - 12/05/08 05:09 PM (15 years, 3 months ago) |
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Just checking.... ^_^
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Nibin
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He is not the only "guru" here but he certainly is one of the most active ones so it is easier to find posts by him.
Here are some of the things that make us trust his advice. There are a few other users who also share many of these characteristics
He has been growing edibles and actives for many many years and as a few other select members actually takes his studies quite scientifically.
He knows Stamets personally and actually disagrees with him on some things, which shows he doesn't just take whatever he says as absolute truth, but backs things up with his own experience.
He has grown shrooms on the most random things like a Bra, a bible, weed, etc.
He has contributed things like the shotgun terrarium, the videos etc.
There are a few other old hands which are just as knowledgeable as him, but they aren't as prolific when posting in the normal mush cult forum.
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alkylbenzene23
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Re: LC Cloning vs Isolating [Re: Nibin]
#9374764 - 12/05/08 06:48 PM (15 years, 3 months ago) |
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A... Bra?
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arp180
student of life
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check out RR's Gallery and you will see it.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: arp180]
#9374833 - 12/05/08 06:57 PM (15 years, 3 months ago) |
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Wow... Well, I'm officially a believer. haha
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LightShedder
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Not only have I done LC cloning before and got varied results from the LC, but I have personally, on multiple occasions in fact, seen a piece of flesh (even from LC clones) sector out into as many as 6 substrains on agar.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: LightShedder]
#9374916 - 12/05/08 07:06 PM (15 years, 3 months ago) |
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Okay, a question now...
LC is obviuosly something that even an amateur shoomerite can do (myself for example). Is agar just as simple, or does it actually require more skill and scientific knowledge/equipment?
Also, how exactly is isolation done?
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Stimpy913
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Quote:
alkylbenzene23 said: A... Bra?
And a burrito. I like his videos. If you try to read up on things here, most of the time people blow it way out of proportion with complication and uncessesary things. In RR's video its' just straight facts, and a good demonstration of idiotproof methods. If i never saw the video, and just read the shroomery, there'd be no growing because the videos show you how easy it really is. Plus it's cool to see someone grow oysters & shiitake off cakes. I never knew that could be done before.
-------------------- How to talk to cops
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J3illy
Trainee
Registered: 10/18/08
Posts: 3,344
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Quote:
Hallucinogenist said: LC is obviuosly something that even an amateur shoomerite can do (myself for example). Is agar just as simple, or does it actually require more skill and scientific knowledge/equipment?
Also, how exactly is isolation done?
From reading about both, I'd say working w/ agar is gonna be a step up from just making an LC. Sterility has to be totally on point w/ agar, just cuz that shit is used specifically for growing out bacteria and spores - so any spec of contamination that may be present will flourish. You need sterility in all aspects obviously, but w/ agar it's MAJOR.
And isolation is getting spores/flesh on agar, and once it starts growing, there will be separate sub-strains. You then transfer a small piece of a sub-strain to a new agar dish, and once that's the ONLY one growing - you've isolated it.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
#9375144 - 12/05/08 07:37 PM (15 years, 3 months ago) |
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And how do you identify the sub-strain on the agar? Microscopically, or am I misunderstanding the Agar process?
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J3illy
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A single substrain is like it's own totally differentiated growth. For example in this pic, the top 1 is isolated, but the bottom 1 looks like many sub-strains growing together. You'd transfer a small piece from the bottom 1, and wait to see how it grows, and keep transferring until it's isolated.
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Hallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
#9375203 - 12/05/08 07:47 PM (15 years, 3 months ago) |
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You mean you'd actually FRUIT the piece from the bottom right? To see if it's correct?
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iMtRiPpIn
Lookin' fer Luv
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Quote:
Hallucinogenist said: You mean you'd actually FRUIT the piece from the bottom right? To see if it's correct?
I think he means you would take a piece from the bottom picture, and put it on agar and grow it out, and keep repeating the process until you get growth that looks like the top picture.
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LightShedder
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I use the grocery store agar tek. It is just as easy as LC. I use short half-pint jars (or maybe they are pint I am not sure). The lids are upside down (for easy opening) closed tightly with a 1/4x1/4 inch hole in the middle stuffed with polyfill.
In the grocery store agar tek he calls for "1/2 of a 9g strip" of solid agar. I like to find the powder agar from health stores and just weigh out 4.5 grams. It is easier then breaking up all the strips.
Also make sure if you get the powder agar that it doesn't have suger or something mixed in. You want plain agar agar.
Once you have an agar plate with one substrain on it (like picture 1 above) then you can make multiple agar plates from it to have a good amount of it stored up. Then you can take a tiny chunk of the mycelium from the plate and drop it in a LC. You can turn the LC into 100s more LCs and then have a huge amount of this isolate stored for use. Just make sure to test out the strain before you go about making so much culture so you know if you even want it.
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J3illy
Trainee
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Re: LC Cloning vs Isolating [Re: iMtRiPpIn]
#9375292 - 12/05/08 08:03 PM (15 years, 3 months ago) |
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Quote:
iMtRiPpIn said:
Quote:
Hallucinogenist said: You mean you'd actually FRUIT the piece from the bottom right? To see if it's correct?
I think he means you would take a piece from the bottom picture, and put it on agar and grow it out, and keep repeating the process until you get growth that looks like the top picture.
Exactly. You cut a really small piece of it, and transfer to a new plate. Then once you have the isolated plate, it's like your master copy - all you gotta do is cut a little piece and you can start a new plate, use it for an LC or inoculation. You'd just keep it in the fridge and it'd be good for a while. For really long-term storage, they have these things called "slants", and the isolated culture will last years.
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J3illy
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Re: LC Cloning vs Isolating [Re: LightShedder]
#9375312 - 12/05/08 08:07 PM (15 years, 3 months ago) |
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Quote:
LightShedder said: I use the grocery store agar tek. It is just as easy as LC. I use short half-pint jars (or maybe they are pint I am not sure). The lids are upside down (for easy opening) closed tightly with a 1/4x1/4 inch hole in the middle stuffed with polyfill.
How does everything work out, do you get many contams? I definitely wanna start working w/ agar, but I'm just kinda discouraged about thinking of contams. If I had a flow hood it'd be a different story, lol. Do you do everything in a glove-box?
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arp180
student of life
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Re: LC Cloning vs Isolating [Re: J3illy]
#9375319 - 12/05/08 08:08 PM (15 years, 3 months ago) |
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"From reading about both"
I am glad to see you started your post with this. I am just starting to read about agar and isolation work. The description and pictures you cited were very helpful. Thanks J3illy
Edited by arp180 (12/05/08 08:10 PM)
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iMtRiPpIn
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Re: LC Cloning vs Isolating [Re: arp180]
#9375349 - 12/05/08 08:13 PM (15 years, 3 months ago) |
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Quote:
arp180 said: "From reading about both"
I am glad to see you started your post with this. I am just starting to read about agar and isolation work. The description and pictures you cited were very helpful. Thanks J3illy
yeah thanks J3illy the pictures were defintiely worth a thousand words. my head starts spinning when i hear terms like "sectoring" and shit but seeing the beginning and end result spoke volumes.
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nw_shroomy
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Re: LC Cloning vs Isolating [Re: arp180]
#9375366 - 12/05/08 08:15 PM (15 years, 3 months ago) |
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I'm sure this can be answered with search but since its being discussed. How long after transferring to agar does it take to grow before its time to transfer to the next agar?
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LightShedder
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Re: LC Cloning vs Isolating [Re: J3illy]
#9375367 - 12/05/08 08:16 PM (15 years, 3 months ago) |
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Yeah I do everything in a sealed still-air glovebox. The only way to pretty much ensure 0% contaminations everytime IMO.
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J3illy
Trainee
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Re: LC Cloning vs Isolating [Re: arp180]
#9375370 - 12/05/08 08:16 PM (15 years, 3 months ago) |
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Quote:
arp180 said: "From reading about both"
I am glad to see you started your post with this.
That's the tone that my posts pretty much always have. I don't claim to have done much yet, but I sure have read a hell of a lot - I mean, look at my title I've had since I joined lol.
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J3illy
Trainee
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Re: LC Cloning vs Isolating [Re: nw_shroomy]
#9375380 - 12/05/08 08:19 PM (15 years, 3 months ago) |
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Quote:
nw_shroomy said: I'm sure this can be answered with search but since its being discussed. How long after transferring to agar does it take to grow before its time to transfer to the next agar?
I think it would take like a week to have the whole plate fill up. But you'd be seeing growth in a few days
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LightShedder
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Re: LC Cloning vs Isolating [Re: nw_shroomy]
#9375381 - 12/05/08 08:19 PM (15 years, 3 months ago) |
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anywhere from 3-7 days depending on that particular substrain and temperature etc. Usually the chunk will recover after 2-3 days and grow from there. I like to do the isolations as soon as you see any growth coming from the chunk. This is to minimize contamination and also to prevent the further mating of new strains.
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nw_shroomy
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Re: LC Cloning vs Isolating [Re: LightShedder]
#9375514 - 12/05/08 08:40 PM (15 years, 3 months ago) |
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Quote:
LightShedder said: anywhere from 3-7 days depending on that particular substrain and temperature etc. Usually the chunk will recover after 2-3 days and grow from there. I like to do the isolations as soon as you see any growth coming from the chunk. This is to minimize contamination and also to prevent the further mating of new strains.
How many times Does it take to get to the one you want?
-------------------- Spawn Ratio Calculator http://www.shroomery.org/forums/showflat.php/Number/7803673#7803673 I only grow edibles.Any info I give ONLY applies to gourmet mushrooms.
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LightShedder
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Registered: 08/30/05
Posts: 3,026
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Re: LC Cloning vs Isolating [Re: nw_shroomy]
#9375567 - 12/05/08 08:45 PM (15 years, 3 months ago) |
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depends. take the smallest chunks possible and you will minimize the amount of transfers necessary but even then, it is dependent on the amount of substrains originally on the plate. It shouldn't take more then 3 or 4 transfers.
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