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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23984795 - 01/05/17 06:32 PM (7 years, 25 days ago)

Quote:

kenetic said:
Quote:

KauaiOrca said:
I've got a killer way of taking prints now that has been contam free for over a year.




Beam me up scotty, I'm down for new methods for taking prints.  I've tried a few different methods and I'm still not sure which one I like the best....




What I do is I make a foil inner liner for a PP5 Pint jar and then cut out a circle smaller than a mushroom cap ... It looks like this ...

From the top...



from the side...



Then I wrap the top in foil and put a kind of foil tent over the whole thing and then PC it for 10 minutes. 

Then, I just take a nice cap, cut it with a red hot knife, place it in the foil liner over the round hole and give it a day.  Then remove the foil liner and all and put the top that was wrapped in foil back on the PP5 Jar.  The cap never physically touched any part of the jar and the jar was sterile.  So the only thing on the bottom of the jar are the spores that were shot out.  I then shoot 10ml sterile water into the jar, swirl it for a few minutes to mix in all the spores and then suck them back up into the syringe ... last two steps in a SAB of course.

Those syringes have been ultra clean  Have put 3 of them on Agar and not even a hint of contam.  Now I skip the agar and just put them on a pint of RGS, do a small grow and that's where I get the mushrooms for the clone water technique.  That way, all the clone water mushrooms are Gen 1 ... and there's no need to ever clean spores on Agar anymore.  I used to get pins from that and put the on Agar, now I just skip that and go clone water to small grain jar.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


Edited by KauaiOrca (01/05/17 06:40 PM)


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OfflineKenetic
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #23984811 - 01/05/17 06:41 PM (7 years, 25 days ago)

Wow man, that's an awesome way to get a clean syringe!  If I wanted to make a print instead I imagine I could put a small square of foil at the bottom of the container and then sterilize it.  Put it in my sab afterward.  Later I could just remove the printed foil from the dish and fold it up. 
I'm not a big fan of syringes but I can easily adapt this method for my use lol.

Awesome!

Edit:  can you think of a way to apply this to a bulk print scenario?


--------------------
Todo Cambia
   

               
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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23984821 - 01/05/17 06:44 PM (7 years, 25 days ago)

Quote:

kenetic said:
Wow man, that's an awesome way to get a clean syringe!  If I wanted to make a print instead I imagine I could put a small square of foil at the bottom of the container and then sterilize it.  Put it in my sab afterward.  Later I could just remove the printed foil from the dish and fold it up. 
I'm not a big fan of syringes but I can easily adapt this method for my use lol.

Awesome!




You can definitely put foil in the bottom ... I've done that too ... When I do that, I just put the unfoil'd top back on, then leave it cracked just a bit and put it in a warm dry place for the inside of it to really dry out ... I use my dehydrator for this ... and then when you open to get the foil it's ultra dry and bacteria free.  I'm telling you, the prints are pristine clean.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23984830 - 01/05/17 06:47 PM (7 years, 25 days ago)

Quote:

kenetic said:

Edit:  can you think of a way to apply this to a bulk print scenario?




Bulk print meaning a bunch of prints at once?  I've never had a need to take more than two at a time ... I can use one of those syringes, because they're so strong, for a lot of jars but what I usually do is just shoot like 3-4 ML's in a grain jar and it colonizes 2-3 times faster ... then I get the mushrooms for clone watering in about a month total or I just use the colonized grain to do a grain water slurry for a new batch of jars.  I've never had contam problems but then again, I don't ever spawn to bulk so that may be why.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


Edited by KauaiOrca (01/05/17 06:49 PM)


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OfflineKenetic
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #23984850 - 01/05/17 06:55 PM (7 years, 25 days ago)

Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23984857 - 01/05/17 06:57 PM (7 years, 25 days ago)

Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.



i think it was wildernessjunkie who posted a great mass printing tek, i just lay foil out in sab


--------------------

It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: cronicr]
    #23984864 - 01/05/17 06:59 PM (7 years, 25 days ago)

Quote:

cronicr said:
Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.



i think it was wildernessjunkie who posted a great mass printing tek, i just lay foil out in sab




The thing is, if the cap physically touches the foil, the chances that some mold and bacteria is transfered to the foil is extremely high....


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23984871 - 01/05/17 07:03 PM (7 years, 25 days ago)

Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.




OK, here's an idea.  If you've got a big PC then just expand the size.  Use a bigger PP5 or even a small pot ... put a much bigger foil liner in it but cut out 5 - 10 small circles in it and put a big piece of foil or a bunch of small ones on the bottom.  You'd have to line it up right and really get those 5-10 caps ready to put inside so nothing floats into the liner ... but I think it's the exact same principle, right?  But you'd still have no caps touching the foil which is where most of the contams are transferred.  I think it would work really good.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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Invisiblewildernessjunkie
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #23985659 - 01/06/17 01:45 AM (7 years, 24 days ago)

Quote:

i think it was wildernessjunkie who posted a great mass printing tek,





Bulk Spore Printing with WJ v2.0

There it is. About 60 prints at once.


Edited by wildernessjunkie (01/06/17 02:32 AM)


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
    #23985698 - 01/06/17 02:23 AM (7 years, 24 days ago)

Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.




Here's another bulk print idea utilizing the same principle.  take a fairly large baking pan that's at least 3-4" deep.  Put your smaller square spore catching foil pieces on the bottom of it.  Then with the heavy style foil, cut a bunch of round holes that the caps will fit over and then stretch the foil over the top of the baking pan so you would essentially have a similar setup.  Then put one more layer of foil over that loosely to cover it up. Then, just put it in the oven to sterilize and since it's metal, you won't have to worry about anything cracking or melting.  When you take it out of the oven, the foil squares on the bottom of the plan are sterile and the piece of foil you'll be putting the caps on is sterile and covered with a protective piece of foil.  Then put that in your SAB and when you've got a bunch of caps, take the protective piece of foil off the top ... put all your caps over the small cut out circles and then leave in in the SAB ... I think that would work really good if I wanted to print like 10-20 caps at a time because, once again, no mushroom caps actually touch the foil that the spores are falling onto which is where 99% of contaminants are transferred to the foil.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


Edited by KauaiOrca (01/06/17 03:44 AM)


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InvisiblefahtsterM
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic] * 1
    #23985791 - 01/06/17 04:25 AM (7 years, 24 days ago)

Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.




Not sure if this relevant to what you're saying here, but I once tried to grow inside an oven bag where I sucked out stale air through polyfil filled tube hosing filters and blow it back in with a fish tank bubbler. then I was going to print into a jar I had in the bag from the beginning..  the idea is that the spores never see unfiltered air and if you print a bunch of caps onto the jar sides and bottom and then seal it up with a silicone coat nut then inject through a SHIP in the jar lid and use a magnet to pull the nut against the sides of the jar through the glass to release them into the water to be sucked back up... clean as a whistle.

Here I'll get the link.. I actually did get the bowl to fruit but the fuckers were sporeless. :mad:

https://www.shroomery.org/forums/showflat.php/Number/7656684

It was overkill, but a fun experiment.. wish they wouldnt have come out sporeless though.

Faht


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: fahtster]
    #23985801 - 01/06/17 04:38 AM (7 years, 24 days ago)

Quote:

fahtster said:
Quote:

kenetic said:
Yeah I know bulk isn't your thing, I was just hoping you could brainstorm some ideas of printing a bunch of caps at once.  If you're smart enough to come up with this, I'm sure you can think of something lol

It's cool though.  Thanks for the info.




Not sure if this relevant to what you're saying here, but I once tried to grow inside an oven bag where I sucked out stale air through polyfil filled tube hosing filters and blow it back in with a fish tank bubbler. then I was going to print into a jar I had in the bag from the beginning..  the idea is that the spores never see unfiltered air and if you print a bunch of caps onto the jar sides and bottom and then seal it up with a silicone coat nut then inject through a SHIP in the jar lid and use a magnet to pull the nut against the sides of the jar through the glass to release them into the water to be sucked back up... clean as a whistle.

Here I'll get the link.. I actually did get the bowl to fruit but the fuckers were sporeless. :mad:

https://www.shroomery.org/forums/showflat.php/Number/7656684

It was overkill, but a fun experiment.. wish they wouldnt have come out sporeless though.

Faht




I'm convinced the contamination that gets on a print (the foil, for instance) comes from the cap touching the foil to make the print.  That's a much bigger risk than the air which can be fairly easy mitigated with a SAB.  My prints got "clean" when I figured out a way to get the spores onto the foil without the cap touching the foil at any point.  By suspending the cap above the foil that catches the spores, that takes the biggest source of contamination AWAY from the print itself.  And it really works.  Add to that, the foil print itself is sterile to begin with and you have a very high likelihood of a very clean print.  Then, if the cap is grown invitro, I think the chance of contamination is probably <1% if that.  But even if the cap isn't invitro, I'm finding virtually no contamination from prints done this way.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #23985806 - 01/06/17 04:45 AM (7 years, 24 days ago)

Oh yeah.. I meant to say sweet idea, to you btw. 

Faht


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Re: Anyone Tried Cloning This Way??? [Re: fahtster]
    #23985814 - 01/06/17 04:59 AM (7 years, 24 days ago)

Screw doing it the hard way. Get prints form lazy basterd tek. No need to care if its perfect since agar work is a must anyway. Even in a trich filled tub i find swabing right under the fruit gets me clean myc on agar to each there own. But having a 100 prints in 100 zip lock bags is hella easy.


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: tump]
    #23985877 - 01/06/17 07:05 AM (7 years, 24 days ago)

Quote:

tump said:
Screw doing it the hard way. Get prints form lazy basterd tek. No need to care if its perfect since agar work is a must anyway. Even in a trich filled tub i find swabing right under the fruit gets me clean myc on agar to each there own. But having a 100 prints in 100 zip lock bags is hella easy.




Taking a clean print and going straight to a small grain grow is a whole lot easier than a dirty print + 3-4 agar transfers to clean it up.  Start it clean and it creates a lot less re-work.  I've got pins showing up for cloning by the time I get a dirty print cleaned up on agar.

I've never understood, given the hyper effort at sterility in this hobby, how being careless with taking the print makes any sense at all.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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Offlinetump
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #23986113 - 01/06/17 09:45 AM (7 years, 24 days ago)

Because no spore print is 100 clean to begin with. With out the three or four transfer on agar i see no point in moving around grain spawn. If we could get spore prints already 100% or even 99.9999% clean we would recondment the spores to lc. I see your point but still hell easy to do in a zip lock


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: tump]
    #24012280 - 01/15/17 01:57 PM (7 years, 15 days ago)

Final update on Clone Water Experiement

Thus far, I've taken clone water from inside a total of 6 mushrooms that was transferred to a small grain jar (pastyplate size).  Here's the final recap of what I learned:

- All 6 small grain jars grew out clean.  Some faster than others, but none of them contaminated in a visible way. 

- When getting the clone water, it's much easier if you pick a thick, half grown specimen with a meaty bottom.  Only need 1-2 ml's of water in the syringe.  You just need to suck back a few drops and it will work.  Even the ones where I wasn't sure I'd even gotten any water sucked back into the syringe ended up growing out fine. 

- I tried dropping the colonized small pucks directly into grain jars but found they colonized much faster if I dropped them in a sterile jar with about 60 ml's of water, shook it to turn it into very thick grain water, then transferred that to the new jars.  Colonization was much faster (3-5 days) this way.

- Varying yields with each jar so far from fantastic to average, but all acceptable.  I suppose this would be true with any cloning method. 

Bottom line is this is a very easy way to clone a growing harvest with very little chance of contamination and no agar step at all.  I didn't even use an SAB to get the clone water and inoculate the small PP5 jar.  Can't say if this is a good process for big spawn to bulk grows, because it probably isn't (too risky) but if you want to grow/test out a bunch of clone cultures with very little fuss, low contamination risk, no agar and the only tool you need is a sterile syringe, it works.  It's a way to go from harvest to next grow over and over because even if the harvest jar was a bit contaminated, the inside of the fruits it produces aren't so the clone water from each harvest will work.  It's quicker and easier than a spore print, a pin to agar grow or a tissue of clone to agar and, I think, opens up the potential of cloning to just about anyone.  All you need is a small PP5, foil, RGS, Syringe, Water, Pressure Cooker and could probably pull it off without the pressure cooker.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


Edited by KauaiOrca (01/15/17 02:21 PM)


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InvisibleAndyHinton


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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #24013887 - 01/16/17 07:17 AM (7 years, 14 days ago)

Update: Reached 50 posts.


--------------------


Edited by AndyHinton (01/26/17 09:37 AM)


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InvisibleKauaiOrca
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Re: Anyone Tried Cloning This Way??? [Re: AndyHinton]
    #24013983 - 01/16/17 08:19 AM (7 years, 14 days ago)

Quote:

AndyHinton said:

Your clone water could inoculate a small grain master, and that culture grown out would be suitable for bulk inoculation. It's another step, but I also work in open air and would be paranoid about inoculating a supplemented substrate with only 3 mL of dirty tissue water.





I'm not convinced at all that the clone water from inside the mushroom is dirty.  In fact, it's proving to be consistently contam free.  Just like working with Agar, there's a certain amount of technique involved to transfers, minimal hand movements, taking the right transfer tissue from the right spot on the plate, etc.  Extracting the clone water is similar.  But if done right, I think the small clone tissue water extracted is very clean, possibly even sterile.  The samples I tested on agar from the small grain jars all looked contam free.

The real purpose of this method is to quickly test a number of half grown fruits (clones) to see if they fruit well ... And it's quite a bit faster than a pin to agar > 2-3 transfers > grow it out to see if it's good > then grow out the master and spawn to bulk ... At least to me, that's pretty time consuming rather than extract the clone water > inoculate small grain jar and grow it out.

The other use for this technique is to simply go from small grow to small grow endlessly without agar or new spores and it's essentially a 15 second technique when you harvest to get another few jars going with clean clone tissue.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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InvisibleAndyHinton


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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
    #24014169 - 01/16/17 09:44 AM (7 years, 14 days ago)

Update: Reached 50 posts.


--------------------


Edited by AndyHinton (01/26/17 09:36 AM)


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