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Invisiblecreamcorn
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LC Cloning - Newbie friendly pictorial tek! * 32
    #5874305 - 07/18/06 07:31 PM (12 years, 9 months ago)

Cloning with liquid culture, the easy way.

This is a tek on how to clone without the need for advanced methods, agar, gloveboxes, flowhoods, or even a pressure cooker.  Its simple enough for a beginning pf-tek'er to complete.  You'll probably also find some good LC tips here if you've been struggling with those.  As always, comments, questions, suggestions, and kudos appreciated! :smile:  I apologize if this is a little wordy, but I wanted to cover all the bases.  I've been answering newbie questions long enough to anticipate them before they've even been asked, so hopefully I got everything covered.

Materials needed:


Step 1.  Prepare a small liquid culture.

For this, I am of the thinking that SIMPLE IS BEST.  My liquid culture containers are simple jars, with a regular old metal lid and ring, with a single hole punched at the top.  The hole should be big enough to accomodate your needle, with a little room to spare so that a vaccum is not created later when aspirating solution.  The hole is covered with a piece of micropore tape, and then covered again either with a coffee filter and rubber band, or a piece of aluminum foil loosely crumpled, to act as a dust cover.


Jar with just a hole

Recipe:  Recipe is the same as any liquid culture; 4% media by weight.  You may use any of the standard LC media, honey, caro, dex/malt will all work.  Water quality is important to LC.  I find that distilled works best, and gives the clearest solution.  You may also use bottled spring water, or tap water filtered with a Brita pitcher or the like.  (Your mileage may vary depending on the quality of your locality's tap water.  In my area, I avoid it.)  Honey quality also varies widely - I've found this particular variety works fantastic for me, and doesn't make any sediment in my LC's even without filtering.  You may need to experiment with brands and types of honey to find one that works.  For a sure thing, you can definitely use karo, and if you're industrious and willing to hunt down dextrose/malt, by all means use that.  The honey I've got seems to outperform karo, so it works for me.


Measure 4 grams of media per 100ml of water

When cloning, I generally start with a small half pint container.  You'll see I've measured exactly 4 grams of honey.  Water weighs 1 gram per 1ml, so I'll fill it to the 100ml point.  I nuke this in the microwave for about 30 seconds, just to warm the water, allowing the honey to dissolve easier, and I stir it up.  Obviously, when making larger LC's, just do the math to stay at that 4% mark.

This will be my "master" culture.  From this culture, small samples are taken to start larger liquid cultures in quart jars that are my "working" culture that I use for inoculation.  The remainder of the master culture can be refrigerated, where it will keep for several months.  When I run out of working culture, it only takes a few drops of the master to start a whole quart of working culture all over again.  Its important to take from the master each time, rather than go from working culture to working culture, so that each of your working cultures are "second generation."

Step 2.  Sterilization.

In my opinion there's only two acceptable ways to sterilize LC for consistent results: steaming, and pressure cooking.  This gives those an option who don't own a PC.  Microwaving works for some, but I don't trust it.  If it works for you, fantastic, go with it.

In both cases, wrap the top of your jar in tinfoil as usual, and take a syringe about half full of water, and also wrap it in tinfoil.  We'll sterilize them both at the same time.


Syringe with some water; wrapped in foil and ready to sterilize

Steam sterilization:
Place a cloth at the bottom of a pot with a tight fitting lid.  Use an inch or two of water.  Place your jar and syringe inside.  Bring to a slow rolling boil.  Once a boil has been achieved, start your timer for 30 minutes.  At the end of 30 minutes, turn off the heat, walk away and don't even think about it for a few hours.  Let it cool.

PC sterilization:
Use your PC as per its instructions with something to keep the items off the bottom of the pot (like a cloth, or a rack if you got one.)  Slowly bring up to pressure.  10psi is plenty if that's all you got.  15psi is fine too.  Do not exceed 15psi.  Once at desired pressure, set your timer for 15 minutes.  DO NOT exceed 15 minutes.  When the time is up, cut your heat and walk away until its cooled.

Step 3.  Select specimen.

Find a nice looking fruit!  We want to clone desirable qualities.  Many have different ideas as to which fruit to clone; ie. choosing the first one up might give you a speedy isolate to work with later, choosing a big one might give you a genetic predisposition towards large fruits; and so on.  I believe that there's just so much variation in growing methods, even though we are taking a clone, we have no idea how closely its going to resemble its "parent" when grown out later - so I take a nice looking fruit and hope for the best.  You'll find out in time if it was a good choice or not when you get your first crop of clones in, and can assess how long they took to grow, the yields you got, and of course, the potency after sampling some.  If it didn't turn out as good as you were hoping, don't fret, you can try again from a different specimen from a different multispore grow.  If it did turn out for you, treat that master culture like gold!

Step 4.  The biopsy

Here's where you'll want to practice your sterile procedures.  No need to go overboard, all I do is ensure there's no drafts, clean my work surface, wash my hands with antibacterial soap, and hit them a second time with some hand sanitizer.  If spraying Oust into the air and washing everything with bleach or whatever makes you happy, knock yourself out. 

Take your fruit, and clean the stem thoroughly with an isopropyl soaked cotton swab.  Clean it all the way around.  Do not touch any areas with your hand near the site of the biopsy.  Quickly wipe your needle with iso (flame if desired, I don't bother - we did just sterilize it after all!) and stab it straight through the stem to the other side.


Swabbing the stem; taking the biopsy.  (This is where 3 hands would come in handy, somebody had to work the camera!  You'll obviously want to hold the mushroom near the top and swab the bottom, and when taking a biopsy not allowing the mushroom to touch any surfaces.)

Remove the foil from your prepared LC jar.  Using a dry cotton swab, clean up any condensation/moisture on the lid.  Then using an alcohol soaked one, clean the lid again.  Give the needle another wipe, insert into hole, and push the plunger.  The water in the syringe will force the cross section of the stem out into your LC.  Quickly cover the hole with a piece of micropore tape (if your lid is still wet with alcohol, that's fine, the tape will stick just fine once it dries) and cover with a dust cover of your choice.


Knock 'er up!  You might need to push a little hard, but the liquid will force the piece of stem out.


You can sort of make out the piece of stem in there.  My solution really was clear I promise, this was without a flash to reduce glare and hopefully get a picture of the piece inside.  On the right, foil loosely back on as a dust cover, don't forget to label!


She's looking a little beat up after being stabbed!

Incubate, and wait.

7-10 days later you should have a nice looking cloud of mycelium.

As they do on the cooking show.... "here's one we've prepared earlier!"  When doing all the above steps, you'll want to work quickly to avoid contamination.  I obviously stopped to photograph everything so who knows if we got a clean sample here.

Here's a "master" culture, about 10 days colonizing.  You'll notice some is already missing, as I've used some of the solution to inoculate larger LCs.


The master culture.  Notice its very thick!  Obviously my little micropore'd hole provides all the gas exchange you need.  You'll also note a few dark specs, that's leftover pieces of the stem.  It kind of dissolves and breaks up to an extent.  It turns dark blue/grey, because you bruised it for sure when poking it with a needle!

Here's a working culture, this is 5 days old.  It was started with about 4cc of solution from a master, and then constantly agitated on a magnetic stir plate.  Use it (or make it) if you got it, but its not necessary.  They'll colonize great without, it just takes a little longer.  LC to LC is FAST no matter what.


Enough LC to kill a small army, from a few CC's of the master.

Some additional LC tips:
-Follow the measurements carefully.  You'll get no growth or poor growth if you use too much media.  When in doubt, use a little less than 4%.
-If your myc cloud is thick and can't be sucked up easily, suck up what you can, and squirt it back down into the cloud.  It will break right up.
-To aspirate from my single-hole jars, I just tilt them on their sides, being very careful not to spill any out the hole.  The needle reaches in just fine.  I gently swirl it around and capture as much mycelium in the syringe as possible.  The thicker the better!
-People often ask, "when is my LC done?"  Well, its done when you want to use it!  If there's enough mycelium floating around that you can capture a bunch in your syringe, go for it.  The rest will continue to grow until the nutrients have run out.  When it doesn't seem to be growing any more, you can refrigerate it, and it lasts many months.  (6 or more.)
-When I aspirate, I take an alcohol soaked cotton swab, and swab right over the piece of micropore tape.  I then stick the needle right through the tape, and fill up my syringe.  I quickly cap the syringe, and quickly replace the soggy piece of micropore tape with a fresh piece.  You can certainly use any number of the fancy LC container teks out there, but it doesn't get much simpler than a hole with a piece of tape on it kids.
-LABEL EVERYTHING.  Keep track of dates, strains, generation number, anything you feel relevant.
-A perfectly prepared karo or honey LC should be nearly crystal clear.  Dex/malt will produce a yellowish solution.  If your karo or honey solution turns yellow, you may have overcooked.  Did you follow my sterilization directions to the T?  Either way, a little overcooked will still work, its not the end of the world.  Likewise, sediment from honey, or from malt/dex will not harm a thing - it just makes spotting possible contamination a little more difficult.  Some take some pretty extreme measures to filter their solutions before sterilizing to get the sediments out, I'll leave it up to you.
-Unless you want cracked jars, heed my advice and allow them to cool gradually and naturally.

Quick Cloning FAQ:

Q: Why clone?
A: Cloning ensures the most efficient use of your substrate, the best possible yields, and once you've found a clone that has a desired potency, the most consistent potency!  See, with a multispore inoculation, you end up with many many strains (each dikaryotic spore mating is technically its own strain).  These strains sometimes grow side by side, and sometimes mate a second time by a process known as anastomosis.  So in the end, you've got a cournicopia of genetic materials and strains.  Not all of these are capable of fruiting!  I think of these strains as "guy on the couch" strains - they'll gladly eat your food and drink your water, but never produce any fruits for you.  By cloning an actual mushroom, we're dealing with one single isolated strain that's already proven itself as being able to fruit.  Another benefit to cloning is nice even pinsets.  When you're dealing with all the mixed strains of an MS inoculation, some parts of your substrate are going to be ready to pin before others.  With clones, they're all going to be ready at nearly the same time, resulting in even, consistent pinsets, and more defined flushes.

Q: If I clone a large fruit, will the resulting fruits grown from it be large?
A: Maybe!  If you have identical twin humans and one eats nothing but cheeseburgers and plays Nintendo all day, while the other eats a balanced diet and exercises regularly, are they going to be the same shape and size?  Of course not.  There's genetic predisposition of course, a large fruit obviously has the genetics needed to be capable of producing more large fruits - but conditions have to be right.  Its nature AND nurture.  Nurture is very, very important, so take care of your babies.

Q: If I clone a mutant, will the resulting fruits be mutants?
A: Again, maybe!  It depends on why the mutation occurred.  If there was a genetic problem that caused the mutation, then sure, it will carry over.  You might not end up with the same exact mutations later, but you'll end up with some funky fruits.  If the mutation occurred because the fruit came in contact with a harmful chemical while growing, that's not going to carry over - the result may very well be a healthy clone.  Nature versus nurture again.  Using the human analogy, if I cloned a person with 6 toes (yeah it happens), the resulting clone will be born with 6 toes.  If I clone a human who broke their leg, of course the baby isn't going to be born with a broken leg!  Got it?

Q: Where should I take my biopsy?
A: I've found that near the base of the stem works best.  I can't tell you why, I don't know, just my experience.

Q: How mature should the mushroom be before cloning?
A: As the viel breaks, or just before.  If it starts to sporulate, it will drop spores on the stem.  You're likely to get spores mixed in with your biopsy, and guess what - its no longer going to be a clone!  You want to avoid spores.

That's all for now, if anybody has any ideas for the little mini clone FAQ above, ask away, and I'll answer to the best of my knowledge (or maybe others can chime in and help out.)  The only way I can make this any simpler is if I were to come to your house and do it for you - and unless you got boobies, and plan to pay travel expenses, it ain't happening. :smile:  Thanks for reading, and good luck.


Edited by creamcorn (07/18/06 08:51 PM)


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OfflineOatman2000
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5874429 - 07/18/06 08:07 PM (12 years, 9 months ago)

I like it!

straight and to the point... having a Magnetic stirer when making lc's, is prolly the best thing i have bought for this hobby... i have 4 different strains of master lc on hand at all times....

sticking the fruit is a very clean way of cloning.... no contact with human hands..

as for the micopore tape for your injection port.. not a big fan of saturating the micopore tape. I go with AGAR's idea of using a Self Healing Injector Port , and a piece of winshield washer hose from auto zone as the filter... they are cheap as hell and no worries of saturation of your filter material.


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InvisibleRoadkillM
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5874447 - 07/18/06 08:11 PM (12 years, 9 months ago)

Nice write up!~

:thumbup: :thumbup:



:popcorn: <--waiting for a newb to ask what an LC is.


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Invisiblecreamcorn
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Oatman2000]
    #5874454 - 07/18/06 08:12 PM (12 years, 9 months ago)

my tape never gets wet with LC.  jars aren't filled to the brim for that reason.  and like i mentioned, i replace the piece of tape with a fresh one anyway each time i draw out solution.  i've done countless LC's and only had the "privlege" of seeing a contamination once or twice.

i'm sure self healing ports and air filters and all that jazz do work spectacular, but like i said i'm a fan of keeping it simple. :smile:  i encourage everyone to do it however it makes them comfortable!


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InvisibleOmnicracker
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Oatman2000]
    #5874477 - 07/18/06 08:17 PM (12 years, 9 months ago)

excellent! i love to see a proper write up.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Omnicracker]
    #5876068 - 07/19/06 01:57 AM (12 years, 9 months ago)

Awesome CC! This is the best LC writeup I've seen, BY FAR! I was going to compliment you on several specific points you make, but there's too many! Every paragraph has at least one nugget of gold in it!

That said, I have some suggestions, which I offer because I think this should become THE LC tek of choice.

1. I would split it into two sections. One for the step-by-step procedure and another for discussion, alternatives, suggestions, and Q&A. That might make it easier for noobs to follow when they're actually performing the procedure. Easier to print the procedure also.

2. As far as 4% by weight, that's awesome that you point this out. I hate seeing "a dab" or "a squirt" that's a recipe for failure if you're not familiar with the procedure.

I would point out that the optimal range is 2-4%, you might mention that. Also honey and karo are partly water, so that should be accounted for if you want to be precise. Here are some values I have in my notes...

Honey = 17.6% water
corn syrup = 19.7% water
High fructose corn syrup = 29% water

Using these figures, with some rounding, it takes about 5g (or ml) of karo to equal 4%. Almost the same for honey.

3. You have karo spelled "caro" in one place.

4. You might mention that dextrose is corn sugar and found in most grocery stores. Also that malt is "spray malt extract" and is available wherever brewing supplies are sold. And that "light" or "extra light" malt extract is best for clarity.

5. You can also boil an LC rather than steam it. I usually boil water and then add ingredients.

6. As far as 15 minutes sterilization it's better to exceed that than go under it. The 10 psi equivalent to 15@15 is 22.5 min.

7. "Enough LC to kill a small army, from a few CC's of the master." Is maybe not the best line, what with search engines and bioterrorism scares and all.

8. You should mention to add the stirbar before PCing. If you don't have a stirrer you should add some sort of agitator, like a crushed marble or shard of glass.

9. I would change "strain" to "sub-strain". I think strain implies more of a genetic difference than you are referring to.

10. You might want to add a link to "FastFred's Media Cookbook"...
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/3077006


Excellent writeup, it's the best one I've seen so far. Make sure to submit it to the FAQ submission forum.


-FF


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InvisibleTippinthru
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5876131 - 07/19/06 02:16 AM (12 years, 9 months ago)


Simple is GOOD. :wink:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5876133 - 07/19/06 02:17 AM (12 years, 9 months ago)

just bookmarked this bad boy. Great write up CC!! 5 for you..


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Yamidude]
    #5876416 - 07/19/06 04:58 AM (12 years, 9 months ago)

Hell yes,I picked a fruit tonight that I will do this with tomarrow.Thanks Creamcorn


Edited by Blutjager (10/08/06 05:39 AM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5876521 - 07/19/06 06:19 AM (12 years, 9 months ago)

Creamcorn, I dont think I've seen you post anything less than gold. This mini tek is no different. I'll be sure to try this method my next flush. The last one I did I had to some dissection to get my stem matter, I never even thought of stabbing the stem with a syringe. Brilliant! Your right, you can anticipate what us noobs are gonna ask before we do lol.


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Invisiblecreamcorn
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5876889 - 07/19/06 10:07 AM (12 years, 9 months ago)

>> That said, I have some suggestions, which I offer because I think this should become THE LC tek of choice.

I never intended on it being an LC tek, it was more or less supposed to be a cloning tek for the LC-familiar, I just got carried away with the LC advice.  I can definitely make a few changes to make the LC aspects more organized/complete and it can serve as both then.

>> 1. I would split it into two sections.  One for the step-by-step procedure and another for discussion, alternatives, suggestions, and Q&A.  That might make it easier for noobs to follow when they're actually performing the procedure.  Easier to print the procedure also.

Will see what I can do...

>> I would point out that the optimal range is 2-4%, you might mention that.  Also honey and karo are partly water, so that should be accounted for if you want to be precise.  Here are some values I have in my notes...

Honey = 17.6% water
corn syrup = 19.7% water
High fructose corn syrup = 29% water

Yep, that's totally right that 4% of corn syrup/honey translates to less than 4% sugars... but like you also say, 2-4% is good, so that lands us somewhere in between.  Its always a safe bet then, and while precise is important, I think simplicity is too.  The "4g per 100ml" rule is just a simple one to follow, and works with any of the media.  (Except high fructose corn syrup... I was under the impression that didn't work so well?  Have never used it to know from experience.)

>> 3. You have karo spelled "caro" in one place.

Oops. :smile:

>> 4. You might mention that dextrose is corn sugar and found in most grocery stores.  Also that malt is "spray malt extract" and is available wherever brewing supplies are sold.  And that "light" or "extra light" malt extract is best for clarity.

Will mention.  I don't work with the dex/malt mix, so my knowledge of the specifics were lacking.

>> 5. You can also boil an LC rather than steam it.  I usually boil water and then add ingredients.

How is boiling then adding ingredients going to give a sterile mixture?  I do mention heating the water to dissolve the media.  Only reason I warm it after to dissolve, is because my scale doesn't have the capacity to measure a jar, with water, and honey. :smile: 

>> 6. As far as 15 minutes sterilization it's better to exceed that than go under it.  The 10 psi equivalent to [Email]15@15[/Email] is 22.5 min.

Again, true.  But when you cut the heat on your PC and let it cool naturally, it still stays at temp (thus still sterilizing) for quite a while.  I have an electric stove, and a large heavy PC, it takes almost 20 minutes just to naturally drop from 15 to 0psi... but I suppose not everyone is in that same boat.  I have personally cooked at 10psi for only 15 minutes without contam... Liquid is easy enough to sterilize, but you can overcook LCs which is whats with the strict time suggestion - but I will edit to increase the time suggestion for 10psi users.

>> 7. "Enough LC to kill a small army, from a few CC's of the master."  Is maybe not the best line, what with search engines and bioterrorism scares and all.

Haha... just a figure of speech but you're probably right given the world we live in nowadays. :smile:

>> 8. You should mention to add the stirbar before PCing.  If you don't have a stirrer you should add some sort of agitator, like a crushed marble or shard of glass.

Yup.  I purposely do not agitate my "master" culture.  That little tiny piece of mushroom flesh gets soggy and breaks up as it is, and its barely viable due to its small size and after all the isopropyl and whatnot, so I leave it undisturbed.  I should probably point that out, as well as a reminder to add your agitators before sterilizing for those who will use them.

>> 9. I would change "strain" to "sub-strain".  I think strain implies more of a genetic difference than you are referring to.

I always thought sub-strain was a bit of a misnomer?  I've honestly never looked up the actual definitions... either way you're probably right again that using the term will make it a little more clear.

>> 10. You might want to add a link to "FastFred's Media Cookbook"...
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/3077006

Will do, hadn't ever previously come across that thread and there's some interesting ideas in it.

>> Excellent writeup, it's the best one I've seen so far.  Make sure to submit it to the FAQ submission forum.

Thanks!

Will get to the editing later today, I shouldn't even be spending this much time here when I should be working as it is. :smile:


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InvisibleTippinthru
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: 2FiNiTe]
    #5876970 - 07/19/06 10:48 AM (12 years, 9 months ago)



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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5877016 - 07/19/06 11:06 AM (12 years, 9 months ago)

> high fructose corn syrup... I was under the impression that didn't work so well?

I've never used it either. I thought it wouldn't work because of some paper I read, but I think it was actually another edible species. I later looked it up in a paper on P. baeocystis and apparently it works for that species, so I'm guessing it would work. It's not very good though. Of the sugars they tested the ranking was maltose, glucose, trehalose, fructose, galactose, sucrose, and lactose. Sucrose barely grew anything and lactose produced nothing.

> Its always a safe bet then, and while precise is important, I think simplicity is too.

I agree. I'm probably too much of a stickler. It's just that I couldn't possibly bring myself to write 4% on media that actually had 3.4%. I would have to bitch slap myself for that, whereas I only curse because of rounding errors.

> How is boiling then adding ingredients going to give a sterile mixture?

I guess I'm too used to making solid media. You have to boil to dissolve agar, and I figured the LC might reach a slightly higher temp with direct boiling over steaming.

> my scale doesn't have the capacity to measure a jar, with water, and honey.

Hmm... I guess it wouldn't work to well to use weighing paper or a boat to measure honey. And I suppose it dissolves easily. Malt is a little tougher. My tips for malt are to use distilled water (for sure), bring that to a low boil, preferably with mag stirring, and *slowly* add the malt *then* the agar (for solid media). Malt sometimes will clump and become a PITA to dissolve. If you make the mistake of mixing malt and dex before adding it's even worse.

> I always thought sub-strain was a bit of a misnomer?

People love to argue that point to no end. If you are of the opinion that Paul Stamets is a god, then you should flame me for blasphemy.

I just disagree with calling each individual a separate strain. Sibling spores share 50% of their DNA and 95% of the time even an expert couldn't determine any phenotypic difference between them. Calling something a separate strain implies a genetic and phenotypic difference. Sibling spores have the absolute minimum of genetic variation possible between two individuals without being clones, so I don't see any way you can call them separate strains. But some people even want to call each sector from the same myc a separate "strain" for Christ's sakes.

You can call them individuals, isolates, cultures, or substrains.

> I never intended on it being an LC tek

Just point out that you can also inject spores and it'll be a great LC tek.

Mad props for the kick-ass write-up.


-FF


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5877036 - 07/19/06 11:17 AM (12 years, 9 months ago)

Quote:

fastfred said:
> How is boiling then adding ingredients going to give a sterile mixture?

I guess I'm too used to making solid media.  You have to boil to dissolve agar, and I figured the LC might reach a slightly higher temp with direct boiling over steaming.





haha reminds me of working with the original homestead kit wayyyys back... getting it home, skimming the instructions... PC?  who needs a PC?  i'm gonna cook up this "agar stuff" in a pickle jar directly on the stove!  which... worked. :smile:  but then i got stuck with it was time to PC their little test tubes of grain, so i had to give in to "the right way"

i guess i wasnt thinking thru when i responded, i suppose it would work if you heated -> added ingredients -> boiled entire mixture direclty over burner in sealed (foil topped for steam escape) container.  from many many steam sterilized jars though i can garuntee it works with a really high success rate, and is probably safer from a jars-cracking oh-shit-there's-lc-everywhere point of view :smile:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5877067 - 07/19/06 11:40 AM (12 years, 9 months ago)

> safer from a jars-cracking oh-shit-there's-lc-everywhere point of view

Hell yes! Never directly heat a glass jar on a stove! Let me tell you, it's a bitch to clean up a liter of media that's flowed into every crack and then gelled. I thought I was being all smart by heating it slowly on a hotplate/stirrer, then BAM media everywhere!

BTW what strain did you get with your homestead kit?


-FF


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5877075 - 07/19/06 11:44 AM (12 years, 9 months ago)

Quote:

fastfred said:
> safer from a jars-cracking oh-shit-there's-lc-everywhere point of view

Hell yes!  Never directly heat a glass jar on a stove!  Let me tell you, it's a bitch to clean up a liter of media that's flowed into every crack and then gelled.  I thought I was being all smart by heating it slowly on a hotplate/stirrer, then BAM media everywhere!

BTW what strain did you get with your homestead kit?


-FF




There was absolutely nothing denoting a strain anywhere in the package... just a little glassine envelope with half a print that was labelled "These are your spores." :smile:  I read somewhere on here they were amazonian in that original agar/grain/compost kit.... its unfortunately no longer with me, this was many many years ago.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5877154 - 07/19/06 12:32 PM (12 years, 9 months ago)

LOL I had that kit that was 6 years ago.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: EquilibriuM]
    #5879389 - 07/19/06 11:56 PM (12 years, 9 months ago)

What's an LC?

splif


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: splifner180]
    #5879399 - 07/19/06 11:58 PM (12 years, 9 months ago)

Quote:

splifner180 said:
What's an LC?

splif


LepreChaun


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Yamidude]
    #5879604 - 07/20/06 12:55 AM (12 years, 9 months ago)

I think LC stands for Leper Choad. Which are often found to be horribly infected.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5925162 - 08/02/06 01:52 PM (12 years, 8 months ago)

Quote:

fastfred said:I thought I was being all smart by heating it slowly on a hotplate/stirrer, then BAM media everywhere!




Ha! Been there man.

I just followed this last night and though I messed up the ratio a little, I hope it works out. Thanks for a great post!


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Tippinthru]
    #5925995 - 08/02/06 06:36 PM (12 years, 8 months ago)

Quote:

Tippinthru said:

Simple is GOOD. :wink:




I've been lokin into these... .2 micron filter.
My G/F has magazines to order these out of. i love the simple design, plus it's only 1 way.

what's the size of drillbit you use for 1.) Filter 2.) stopper.

"Thanks FF for correcting me"


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Edited by Oatman2000 (08/02/06 07:05 PM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Oatman2000]
    #5926050 - 08/02/06 06:55 PM (12 years, 8 months ago)

That's a .2 micron filter, not .002 micron. And they cost about $5 each. Polyfill is a lot more cost effective.


-FF


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #5927370 - 08/03/06 01:26 AM (12 years, 8 months ago)

Quote:

fastfred said:
That's a .2 micron filter, not .002 micron. And they cost about $5 each. Polyfill is a lot more cost effective.


-FF




http://www.gearboxinc.com/product_info.php?cPath=28&products_id=73

correct. .2 and at least 4.50 each unless you buy bulk


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Yamidude]
    #5927960 - 08/03/06 09:41 AM (12 years, 8 months ago)

well since that all my bad thoughts on LC are now cleared up, I cant wait for my spores... That has helped me alot... made the thought of lc so basic for me (figure of speach)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Delkor]
    #5929760 - 08/03/06 10:52 PM (12 years, 8 months ago)

hey creamy corn licker... I just used this tek in the past week and I'll have to say, I have a footnote to add... it's easier for me to get a nice chunk of myc that fills the needle by inserting the needle in a diagonal direction up the stalk... it works really good, a lot better than straight through... I cloned the 11 inch badboy and it's really taking off... I cloned that one 4 days ago and it's already growing myc off the spaghetti lookin biopsy chunks :laugh: :laugh: :laugh: :laugh:

I love you man, and you have a special place in my boner pants.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: liamtheloser]
    #5929771 - 08/03/06 10:59 PM (12 years, 8 months ago)

You best save me a print once you get some giants if you know whats good for ya..


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Yamidude]
    #5929800 - 08/03/06 11:17 PM (12 years, 8 months ago)

you want the print off my giant? I'll totally send it to you. It's faint (which is why i decided to clone in the first place), but it's yours if you want it

pm me your address and i'll send it off on monday


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: liamtheloser]
    #5929862 - 08/03/06 11:44 PM (12 years, 8 months ago)

Quote:

liamtheloser said:
hey creamy corn licker... I just used this tek in the past week and I'll have to say, I have a footnote to add... it's easier for me to get a nice chunk of myc that fills the needle by inserting the needle in a diagonal direction up the stalk... it works really good, a lot better than straight through... I cloned the 11 inch badboy and it's really taking off... I cloned that one 4 days ago and it's already growing myc off the spaghetti lookin biopsy chunks :laugh: :laugh: :laugh: :laugh:

I love you man, and you have a special place in my boner pants.




yeah... diagonal is a good plan.  meaty mushrooms are good choices not only because meaty mushrooms are a good thing to clone, but you get more tissue... and larger gauge needles obviously are best too.


and for the record, the half pint i made in the pictures for this thread, is sitting in the bottom of my closet, contaminated with all sorts of bacteria.  it started growing fuzz off the tissue chunk, then came all sorts of dusty looking growth, and now it looks like somebody shot a hot steamy load into my jar when you swirl it around.  i wasted a half pint of perfectly good LC for you punks because i was too busy taking pictures to worry about being sterile  :smile:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5931295 - 08/04/06 02:11 PM (12 years, 8 months ago)

Definitely an awesome write up creamcorn :thumbup:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: _OttO_]
    #5940681 - 08/07/06 02:28 PM (12 years, 8 months ago)

Great work, nice pictorial :thumbup:

Sorry for the stupid question, but i dont understand the biopsy part. You just pass the needle through the stem or what?

btw im a cloning noob


Edited by baltazar (08/07/06 02:29 PM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: baltazar]
    #5940739 - 08/07/06 02:47 PM (12 years, 8 months ago)

This is amazing.

I cant wait to try this!!!


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: baltazar]
    #5942023 - 08/07/06 10:03 PM (12 years, 8 months ago)

Quote:

baltazar said:
Great work, nice pictorial :thumbup:

Sorry for the stupid question, but i dont understand the biopsy part. You just pass the needle through the stem or what?




yes.  you trap a piece of mushroom tissue in the needle, and you eject it into your LC solution.  that's really the whole tek in a sentence. :smile:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #5972139 - 08/17/06 07:40 AM (12 years, 8 months ago)

about 3 months ago i was ordering some spores to pop my shroom growin cherry from microtech( ASSHOLES but i did get my money back) and lookin at some teks... most have missed this one

Just one question is this an actual growin tek or just to replace spoers if not how bout some picks of shrooms

Know its dumb question dont think that the shroomin would actually grow in liquid


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: biohaze]
    #5972146 - 08/17/06 07:49 AM (12 years, 8 months ago)

nm lol just noticed that LC teks are not under the basic teks and it just replaces spores


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Roadkill]
    #5972240 - 08/17/06 09:50 AM (12 years, 8 months ago)

^^^^^^

Quote:

Roadkill said:
:popcorn: <--waiting for a newb to ask what an LC is.




well RK, the wait is over. :smile:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #6065422 - 09/15/06 06:35 PM (12 years, 7 months ago)

Creamcorn you are a god! I got hooked on this hobby through a short video, but that was elementary compared to your techniques. You keep it simple and get results, couldn't ask for any more! CC FTW!  :eek:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: xeallos]
    #6065600 - 09/15/06 07:31 PM (12 years, 7 months ago)

Thanks Creamcorn. That is a nice, straight forward, write up.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #6145003 - 10/08/06 12:28 AM (12 years, 6 months ago)

that's a bookmark right there.
thanks for taking the time, bro.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Jack_Straw]
    #6145755 - 10/08/06 04:50 AM (12 years, 6 months ago)

I just tried this with a spore syringe.I hope it works out.


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OfflineIFEELGOOD
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: hucker696]
    #6166528 - 10/13/06 06:44 PM (12 years, 6 months ago)

haha im a failure this sounded so easy but I stabbed my poor mushroom like 50 times and couldn't get a visible piece in the needle. Maybe for me I should just cut off a few lil pieces and put them in the hole manually ? lol

I'm serious I tried I feel like a failure. Maybe one got enough in it and it will come out Ok...


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: IFEELGOOD]
    #6166600 - 10/13/06 07:04 PM (12 years, 6 months ago)

A few things that help are a larger gauge needle, stabbing at an angle while you go through, and twist as you stab - the sort of angled "sawed off" part of the syringe will help you cut a nice core.  Obviously the thicker/meatier the mushroom, the better.  Its not just you, it can be a little tricky at times... so don't feel like a failure, I've screwed up my fair share of these too. :smile:  Luckily it takes a few cents and a few minutes to make another LC jar so there's not much at stake.

You absolutely can cut a piece out and start your LC that way... the coring/biopsy technique is probably the most difficult way to get a sample, but the easiest way to transport a sterile sample from mushroom to culture.  On the other hand you can cut a piece out easy, but the hard part is getting a sterile piece into a sterile culture since it would involve exposing more mushroom, and opening the lid to your LC container.  If you have a glovebox (or even take a few minutes and throw together a ghetto cardboard-box-saran-wrap glovebox) you can definitely pull it off with a scalpel - slice a mushroom open, cut out a chunk, stab it, open LC and quickly transfer inside, and bam.  :thumbup:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: IFEELGOOD]
    #6166601 - 10/13/06 07:05 PM (12 years, 6 months ago)

Are you sure those cultures are clean creamcorn? They appear to be a bit cloudy IMO. Just wondering if you've tried them?


Edited by Hotnuts (10/13/06 07:07 PM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Hotnuts]
    #6166638 - 10/13/06 07:13 PM (12 years, 6 months ago)

The cultures shown worked great.  They're always under refrigeration so when I took them out to take snapshots, the cold made water condense on the outside of the jars... could be what you're seeing?  Or maybe a combination of camera tricks and thick as hell LC. :smile:

In fact, the "dimple cap" substrain I was just going off about on in that other thread about the strains (and in my avatar picture) is what you see in those two jars of colonized LC.

I actually contaminated my master culture (the half pint) of that after the fact. :frown:  I've still got that quart jar that was pictured on the mag stirrer in my fridge since July... little less than a half of it left... just 'nocced some last week and those WBS jars are almost colonized, so its still hanging in there.

Coincidentally, the one I prepared while taking the pictures never even actually worked... I was a bit less than sterile because I was trying to photograph things. :smile:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #6169129 - 10/14/06 04:11 PM (12 years, 6 months ago)

thanks cream i'll probably have to give it another try.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: IFEELGOOD]
    #6169221 - 10/14/06 04:52 PM (12 years, 6 months ago)

What happens if you clone an abort??


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #6462345 - 01/15/07 04:11 PM (12 years, 3 months ago)

Quote:

creamcorn said:
"Its important to take from the master each time, rather than go from working culture to working culture, so that each of your working cultures are "second generation"."

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

OK, why do you want to use a "second generation" liquid culture to inoculate your jars? Can't you just use your master to inoculate your jars and then when you get low you just make another master? Or is this done only to lessen the risk of contaming the master?


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: coastalite]
    #6462518 - 01/15/07 04:52 PM (12 years, 3 months ago)

Excellent stuff. Thanks Creamcorn.  :thumbup: :thumbup: :thumbup:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #6501036 - 01/27/07 01:54 AM (12 years, 2 months ago)

i tried this w/some koh samui specimens i made 6 lc's from 2 fruit bodys. ive done lcs from spore or myc in the past and it has always worked great, this is the first time i ever tried w/a tissue sample. for some reason the only growth i am getting is like extremely off white and ulike the l.c.s i have made in the past it all kind of settled in the bottom whereas in the past the myc always kind of clouded up the whole thing... do you think this is contamination or what? i was worried about using tissue because of contamination due to tissue decay could that be what i am seeing? the sample in each jar is about the size of 1/2 a grain of rice is that too big?


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fetalscab]
    #6532078 - 02/05/07 01:42 AM (12 years, 2 months ago)

I made 3 LCs the other day, i found that my needle was too small to catch tissue from a fresh stem so i took a tiny abort and sliced the stem into thin 1/4 inch thick disks, Pc'd up some karo water and in a clean environment put the stem cuttings into the jars. Its been about 1 1/2 week and dizam i have good growth in the jars, I also did one with spores but it isnt growing nearly as quick as the fresh tissue. The myc sorta grew like a blob around the cutting at first, roping out in strands 360 degrees around le' tissue, but once i shook the jars up a lil and got the myc broken up it grew much thicker.


Edited by large_dose (02/05/07 01:43 AM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: large_dose]
    #6532087 - 02/05/07 01:46 AM (12 years, 2 months ago)

Quote:

large_dose said:
I made 3 LCs the other day, i found that my needle was too small to catch tissue from a fresh stem so i took a tiny abort and sliced the stem into thin 1/4 inch thick disks, Pc'd up some karo water and in a clean environment put the stem cuttings into the jars. Its been about 1 1/2 week and dizam i have good growth in the jars, I also did one with spores but it isnt growing nearly as quick as the fresh tissue. The myc sorta grew like a blob around the cutting at first, roping out in strands 360 degrees around le' tissue, but once i shook the jars up a lil and got the myc broken up it grew much thicker.




Next time use an active pin not an abort. Pick a pin in the center of a nice cluster. There is an impressive thread from RR with pin-clones on agar.


Yeah.. this thread deserved a bump:thumbup:


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: MajorDick]
    #6532102 - 02/05/07 01:49 AM (12 years, 2 months ago)

my bad, it wasnt an abort, it was activly growing.

and heres fastest LC as of today..




You gotta realize how simple this is, sterile karo water and a clean environment to do the transfer. i heard of some users soaking the tissue in h202 water but i put mine right in and had no probs. Its getting pretty dense, the strain is B+, does everything look ok to you guys? This is my first lc clone attempt


Edited by large_dose (02/05/07 11:51 PM)


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: large_dose]
    #6535125 - 02/05/07 11:52 PM (12 years, 2 months ago)

^ bump - uploaded pics.


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OfflineSulli
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: large_dose]
    #6550816 - 02/10/07 04:13 PM (12 years, 2 months ago)

Hey I started my LC yesterday and this will be day one. Every thing is looking good so far. PICS coming soon.

Also has anybody ever stabed their finger with the needle???
I could not get the top of it and when I did my arms did some kind of kick back and stuck the needle right into my thumb, damn it hurt so bad! Any body ever do that?


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


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InvisibleFellowGrower
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: hucker696]
    #7114562 - 07/01/07 12:21 PM (11 years, 9 months ago)

Man O Man...
WHERE HAVE YOU BEEN ALL MY LIFE!
so tell me...
WHat are your thoughts on this...?
http://www.shroomery.org/forums/showflat.php/Number/7093565#Post7093565
I know you've seen it!
you just wanted me to come find this didn't you! lol.
I know how ya'll are!

Thanks Brah.

-the Schlager-


--------------------
[url=http://files.shroomery.org/files/15-009/514830879-Take_over_this_thread.jpg]


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Invisiblew. murderface
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: FellowGrower]
    #7158059 - 07/10/07 10:38 PM (11 years, 9 months ago)

Liquid Culture right?


--------------------
Eat right stay fit DIE anyways.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: w. murderface]
    #7189474 - 07/17/07 08:32 PM (11 years, 9 months ago)

when you are stiring your LC using the magnetic stir plate...how do you get the magnetic stirrer out??? wouldnt you then have contams? or does none of that matter. im SLIGHTLY confused..


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Offlineshroom1957
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: sproket13]
    #7189837 - 07/17/07 10:05 PM (11 years, 9 months ago)

you put the stirrer in b4 you pc and leave it in until you're done with that LC. I use painted paneling nails....don't rust.


--------------------
Reality is just a way of looking at things that you were taught before you were old enough to know any better! -me

For me the world is weird because it is stupendous,awesome,mysterious,unfathomable;my interest has been to convince you that you must assume responsibility for being here, in this marvelous world,in this marvelous desert,in this marvelous time. I wanted to convince you that you must learn to make every act count, since you are going to be here for only a short while;in fact,too short for witnessing all the marvels of it.
-Don Juan


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OfflinePeace
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #7202698 - 07/20/07 04:05 PM (11 years, 9 months ago)

I have a question... will this Albertsons brand of Clover Honey Grade A work? I'm not sure if its organic or not... it just gives 'honey' as the ingredients on the back. I want to test out doing a 50/50 (karo and honey) but cant start till i know its the right honey :\
Any advice is greatly appreciated.
thanks for the tech!


Edited by Peace (07/20/07 04:13 PM)


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OfflineCaptTrip
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Peace]
    #7202871 - 07/20/07 04:53 PM (11 years, 9 months ago)

Quote:

Peace said:
I have a question... will this Albertsons brand of Clover Honey Grade A work? I'm not sure if its organic or not... it just gives 'honey' as the ingredients on the back. I want to test out doing a 50/50 (karo and honey) but cant start till i know its the right honey :\
Any advice is greatly appreciated.
thanks for the tech!





I've tried a couple different brands of honey and haven't noticed much of a difference.

I think it suggests "organic" honey because it is a little clearer, which helps your LC stay clearer.


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


:mushroomgrow: Cambodian Grow Log :mushroomgrow:


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InvisibleBlutjager
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: CaptTrip]
    #7203423 - 07/20/07 07:31 PM (11 years, 9 months ago)

As long as the bottle is shaped like a bear it will work:dielaughing:

No for real,that all I ever use:thumbup:


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OfflinePeace
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #7205369 - 07/21/07 05:17 AM (11 years, 9 months ago)

haha k thanks guys. I'll try doing a Large jar (Quart) with just the normal karo light, and inject it with 3-4ml of Puerto Ricans. Then in another Quart jar do the same but with Ecuador. Then with two half pint jars i will do a 50/50 mix (honey/karo) and the other with straight Honey and inject both with the same strain of either PR, Equador, Golden Teacher or B+... what ya think i should pick? First time growing successfully (got a P.Cooker finally) so not sure which would be best to test for the smaller jars.


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InvisibleFellowGrower
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Peace]
    #7205525 - 07/21/07 07:59 AM (11 years, 9 months ago)

I use Shards from the glass pile I have from bustin up jars, b/c as we have already said... ANY jar will work...
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7136379/an/0/page/0

So, in this sense, you should have a bunch of sterile shattered glass anyway! right? lol

heres what mine looks like.
it started turnin red?
you see it? in the top left. just below the surface, theres a web of myc floatin with red shit on it. and around the rim, just above the water level, little red specs stuck to the side?
whats that mean?

If its bad news keep it to yourself k?:calledajoke:
lol Im kidden whats the deal?


--------------------
[url=http://files.shroomery.org/files/15-009/514830879-Take_over_this_thread.jpg]


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: FellowGrower]
    #7214087 - 07/23/07 04:22 PM (11 years, 8 months ago)

Just a suggestion but it may be better to take the biopsy from the top of the cap straight through the middle of the stem. That
way you only have to wipe the top and the tissue where the cap
meets the stem is usually meaty.

Just a thought.


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OfflinexMoorhsuMx
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #7345968 - 08/28/07 09:34 PM (11 years, 7 months ago)

Amazing Good job man :smile:


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InvisibleThePyschonaut52
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: xMoorhsuMx]
    #7403863 - 09/13/07 08:07 AM (11 years, 7 months ago)

bump


--------------------
"In god we trust..."


-I guess we're screwed.


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InvisibleFellowGrower
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: ThePyschonaut52]
    #7404756 - 09/13/07 04:58 PM (11 years, 7 months ago)

how do You tell when a PC without a gauge is at 15psi??
thatnks.....


--------------------
[url=http://files.shroomery.org/files/15-009/514830879-Take_over_this_thread.jpg]


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: liamtheloser]
    #7447847 - 09/24/07 04:06 PM (11 years, 6 months ago)

This is the best tek i have read on cloning/LC. Im just waiting for pinning on my first cake (please pin and fruit!!!) and after reading this i will be cloning of my first cake.

I already have done 1 lc culture from spores (have no idea how it worked due to using a shot glas covered with tinfoil, and having to take the foil of and break up the myc with the needle without a glove box) i have a 1/2 pint pf tek already 100% colinesed in about 7 days just waiting to make sure it has a hold in the middle (though im sure it has as i inoced in the middle) then its birth. I just threw the rest out as it would have probably contamed.

This tek is so clear, that the only question that should every be asked is - could u simply inject that segment into a shroom and inoculate say a pf tek jar, or even a grain jar. I think it would work, it seems pretyy stupid though as u could only do it a couple of times then the generations would start to degrade and die. BUt if u got a growth from spores that had many good fruits, u could start a quite a few master cultures from various different fruits with qualities u may want to keep on going, then use another peice to start of more jars while u r waiting on the lc's growing.

5 shrooms for u!!!!

This should definatly be put into the teks and made the tek of choice. I have read probably all of them (as far as i can see from the cultivation section) and this is definatly the best.


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

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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Offlinejuende
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: veda_sticks]
    #7447995 - 09/24/07 04:57 PM (11 years, 6 months ago)

wow how did i miss this. this is one amazing write up
good job cubie!


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InvisibleMonstroniuM
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: juende]
    #7514487 - 10/14/07 03:43 AM (11 years, 6 months ago)

Totally didn't think that the clone jars needed gas exchange. I used the left over spores of my syringes after inoculation. There is def. mycelium in the jars, its just not cloudy/milky. There are little pieces, on the bottom, as well as floating. I take it is because I have had the jar lids screwed all the way down (I dont have a fancy gas exchange system like you, just one hole in the lid for the syringes with tape over it). I have just now after re-reading your post loosened the lid, but is it too late? I started the clones on, I want to say 9/20-23(?).

Thanks in advance!

-Monstro


--------------------
The text and pictures posted by this user are entirely fictional. All posts are for entertainment purposes only and do not necessarily reflect the views or beliefs of this user.


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Invisiblefastfred
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: MonstroniuM]
    #7517455 - 10/14/07 10:56 PM (11 years, 6 months ago)

> Totally didn't think that the clone jars needed gas exchange.

They don't. I don't know where you're getting that from, but LCs need no gas exchange except for the pressure equalization required to prevent the jar from sucking in an entire syringe when you inoculate.

> There is def. mycelium in the jars, its just not cloudy/milky.

You don't want your LC to be cloudy or milky. That is a sign of bacterial or yeast contamination. All you want is to be able to see myc and have the rest of the liquid be clear.

If you do open up your LCs and any contaminant filled air gets inside you will indeed end up with a cloudy, milky, contaminated, useless mess.


-FF


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #7518019 - 10/15/07 01:18 AM (11 years, 6 months ago)

Well thank you for clearing it up for me, I can sleep easier now. I didn't open them up I just unscrewed the cap a wee bit so it wasn't incredibly tight, but have since fixed that. Thank you FF.


--------------------
The text and pictures posted by this user are entirely fictional. All posts are for entertainment purposes only and do not necessarily reflect the views or beliefs of this user.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: creamcorn]
    #7841362 - 01/06/08 10:58 PM (11 years, 3 months ago)

Been doing my research for the past 2/3 weeks. I finally understand what an LC really is. It seems so much easier and more effective than a grain to grain transfer. Quick question, do you use the same amount of LC as you do with a spore Syringe? Thanks for such a HUGE help!


--------------------
Phi Alpha


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: HeadFood]
    #7842252 - 01/07/08 05:42 AM (11 years, 3 months ago)

Use as much as you want, you'll have plenty of it. The only thing to worry about is screwing up the moisture content. If you use too much you'll end up with overly wet substrate. You also don't want a lot of sugars in your substrate as that will encourage contamination when you fruit them.

It's better to use the minimum required, but if you want faster colonization then use a little more and spread it around as much as possible.

Your minimum amount will depend on how thick your myc is in the LC. As long as you get a tiny bit of myc at each innoc site you'll do fine. 1/2cc per hole should be a pretty good amount to start with.


-FF


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Invisiblesmokedout420
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #8027920 - 02/15/08 05:59 PM (11 years, 2 months ago)

this kicks ass thanks for the write up


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Offlinejung
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: smokedout420]
    #8031101 - 02/16/08 02:47 PM (11 years, 2 months ago)

i'm gonna try this with shiitake.

but if you're shooting for exactly 4% with 4grams of honey, wouldn't 96ml of water is used instead of 100?


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Offlinejtbyrd
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: jung]
    #8182409 - 03/23/08 04:22 AM (11 years, 1 month ago)

Is that the lid separately wrapped in foil in the original picture? do you not leave it on the jar and wrap it in foil like you would if you were pcing a pf jar? just a bit confused....thanks for the help.


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Offlineultralight
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: jtbyrd]
    #8182542 - 03/23/08 06:03 AM (11 years, 1 month ago)

Quote:

jtbyrd said:
Is that the lid separately wrapped in foil in the original picture? do you not leave it on the jar and wrap it in foil like you would if you were pcing a pf jar? just a bit confused....thanks for the help.





The pic of the lid alone, was just for reference, (a foil covered lid)... always keep the lid on the jar when sterilizing substrate!.


Edited by ultralight (03/23/08 06:14 AM)


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Invisibletempleton1
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: liamtheloser]
    #8280010 - 04/14/08 12:28 AM (11 years, 12 days ago)

awesome tek, thanks


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OfflineOzzy_shroomer
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: templeton1]
    #8518544 - 06/13/08 07:15 AM (10 years, 10 months ago)

Awsome tek i like it and think it should be brought back up for other to read..

also just a thought going down from the cap into the stem might be a good way to clone wild mushrooms :shrug:


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


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Invisiblechadf36
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Re: LC Cloning - Newbie friendly pictorial tek! *DELETED* [Re: Ozzy_shroomer]
    #8518678 - 06/13/08 09:13 AM (10 years, 10 months ago)

Post deleted by chadf36

Reason for deletion: my name is showing up under google searches



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OfflineTbolg
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: chadf36]
    #8772450 - 08/14/08 11:28 AM (10 years, 8 months ago)

Bad ass tek bro. Will be using this to make a MS LC and then after a harvest a cloning . Thanks for the info.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: fastfred]
    #8909406 - 09/10/08 11:36 PM (10 years, 7 months ago)

Quote:

fastfred said:

I've never used it either.  I thought it wouldn't work because of some paper I read, but I think it was actually another edible species.  I later looked it up in a paper on P. baeocystis and apparently it works for that species, so I'm guessing it would work.  It's not very good though.  Of the sugars they tested the ranking was maltose, glucose, trehalose, fructose, galactose, sucrose, and lactose.  Sucrose barely grew anything and lactose produced nothing.


-FF




Interesting info.. If this is the case, try a simple syrup..

Take some sugar and water, about a 1:1 ratio... up to a 1:2 water:sugar

Bring this to a boil.

Cook it until it turns a faint amber color (this step is important. Chemically, you are breaking the sucrose molecules into one glucose and one fructose molecule).

Now you have a solution of glucose and fructose with the added benefit of being sterile.

This tek should work well, since you have two of the better performing sugars, as listed above.

PS: I'd be wary of honey without a PC... while undiluted, it is so hydroscopic that nothing can grow in it... once you dilute it, many dormant spores could begin to grow... not the least of which are botulism spores.. which honey is notorious for being contaminated with... so i would def avoid it if one does not own a PC..


Edited by FaeFire (09/10/08 11:41 PM)


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Offlinecutemushie
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: Tippinthru]
    #9170605 - 11/02/08 01:02 AM (10 years, 5 months ago)

Quote:

Tippinthru said:

Simple is GOOD. :wink:




How does this lid work?  I am assuming that there's a filter for gaseous exchange.  But when you turn it upside down, does the filter not get wet and subsequently wick in contamination?


--------------------
HAVE: Paneolus cyanescence, Paneolus cambodgiensis, Psilocybe cubensis, Flammulina velutipe (white, and brown Korean strain), Hericium erinaceous, Coprinus atramentarius, Coprinus comatus, Agrocybe aegerita, Pleurotus eryngii, Psilocybe subaeruginosa, Ganoderma lucidum, Pleurotus nebrodensis, Lepista nuda, Agaricus augustus, Pleurotus cystidiosus, Volvariella speciosa.

LOOKING FOR: Those that I don't have.


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OfflineHippieChick
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: cutemushie]
    #9170691 - 11/02/08 01:44 AM (10 years, 5 months ago)

Quote:

cutemushie said:
Quote:

Tippinthru said:

Simple is GOOD. :wink:




How does this lid work?  I am assuming that there's a filter for gaseous exchange.  But when you turn it upside down, does the filter not get wet and subsequently wick in contamination?




Read

http://www.shroomery.org/forums/showflat.php/Number/6214730#Post6214730

Peace,Love and Happiness
:heart: HC :mushroom2:


--------------------
Peace,Love and Happiness
:heart: HC :mushroom2:

Freedoms just another word for nothing left to lose..............

I LUV My Greenhouse
http://www.shroomery.org/forums/showflat.php/Number/5545848#5545848

My First Pans
http://www.shroomery.org/forums/showflat.php/Number/6212058#6212058


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OfflineP.Menace
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: HippieChick]
    #9170696 - 11/02/08 01:46 AM (10 years, 5 months ago)

damn... stop wreck bumping and hijacking OLD threads


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

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.


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OfflineHippieChick
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: P.Menace]
    #9170731 - 11/02/08 02:05 AM (10 years, 5 months ago)

What?


--------------------
Peace,Love and Happiness
:heart: HC :mushroom2:

Freedoms just another word for nothing left to lose..............

I LUV My Greenhouse
http://www.shroomery.org/forums/showflat.php/Number/5545848#5545848

My First Pans
http://www.shroomery.org/forums/showflat.php/Number/6212058#6212058


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OfflineP.Menace
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: HippieChick]
    #9170753 - 11/02/08 02:12 AM (10 years, 5 months ago)

not you.. your cool

I was talking to the n00b... but I have taken it to PM.

Sorry for rebumping the old ass thread again


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

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.


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OfflineHippieChick
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: P.Menace]
    #9170826 - 11/02/08 02:39 AM (10 years, 5 months ago)

Quote:

P.Menace said:
not you.. your cool

I was talking to the n00b... but I have taken it to PM.

Sorry for rebumping the old ass thread again




Woo . Thank God . I was really worried about not being cool anymore. Thanks for putting my mild at ease,lol .

Thanks for taking it to PM to straighten that nOOb out . But can I ask you what
exactly you're straightening out?

This is posted as a "nOOb" friendly TEK that's received over 22,000 views and 89 responses that contains a bunch of great info . Some new person takes the time to do what everyone tells him , search and read , and sees something posted in the third response , asks a question about it and gets straightened out via PM ,LMAO .

Didn't know this was on the list of threads not allowed to be bumped.

Peace,Love and Happiness
:heart: HC :mushroom2:


--------------------
Peace,Love and Happiness
:heart: HC :mushroom2:

Freedoms just another word for nothing left to lose..............

I LUV My Greenhouse
http://www.shroomery.org/forums/showflat.php/Number/5545848#5545848

My First Pans
http://www.shroomery.org/forums/showflat.php/Number/6212058#6212058


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InvisibleFooManM
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: HippieChick]
    #9170904 - 11/02/08 02:56 AM (10 years, 5 months ago)

I gotta agree with HC here. It would be one thing if the bump was a pointless smiley or something, but the poster actually had a relevant question. Although Tippin hasn't posted in FOREVER, a noob probably wouldn't know how to check that.


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OfflineP.Menace
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: FooMan]
    #9170947 - 11/02/08 03:10 AM (10 years, 5 months ago)

Quote:

FooMan said:
I gotta agree with HC here. It would be one thing if the bump was a pointless smiley or something, but the poster actually had a relevant question. Although Tippin hasn't posted in FOREVER, a noob probably wouldn't know how to check that.




theres the situation.
NM i guess im the only one who finds 2+ Year old threads come back to the living.
and thread hijacking... this noob has a history today

dont worry bout me, im just some crazy fuck....


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

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: P.Menace]
    #9170993 - 11/02/08 03:26 AM (10 years, 5 months ago)

i love seeing old threads like this pop up on my thread list
i like looking back on the posts i made years ago,
its funny looking back.

theres quite a few helpful threads id like to see come back to life,
im about to bump them, can i?... if the question is relevant.

usually the teks with quite a few pages written by experienced members are the one's worth reading,
unless its littered with drama of course.
where's creamcorn, anyways?


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: large_dose]
    #9171001 - 11/02/08 03:29 AM (10 years, 5 months ago)

Creamcorns last post was 01/15/08 05:29 PM


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

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.


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Re: LC Cloning - Newbie friendly pictorial tek! [Re: P.Menace]
    #9171003 - 11/02/08 03:30 AM (10 years, 5 months ago)

Solid cloning tek, especially for individuals who lack agar media and petri dishes.  Glad it got bumped,  dunno the reason it got bumped though.


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Sh sh shaw!



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Re: LC Cloning - Newbie friendly pictorial tek! [Re: T2K_Rusty]
    #9171007 - 11/02/08 03:33 AM (10 years, 5 months ago)

wow, informative, i didnt know that i could view a list of his posts...:lol:

jk

i didnt know if anybody else knew him personally, and had a clue what he was up to.


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InvisibleFooManM
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Re: LC Cloning - Newbie friendly pictorial tek! [Re: P.Menace]
    #9171065 - 11/02/08 03:57 AM (10 years, 5 months ago)

Quote:

P.Menace said:
dont worry bout me, im just some crazy fuck....




:lol:


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OfflineAnnoA
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Re: LC Cloning - Newbie friendly pictorial tek! (moved)
    #9171541 - 11/02/08 10:28 AM (10 years, 5 months ago)

This thread was moved from Mushroom Cultivation.

Reason:
-> archive


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