Paradox
©
Fisana

Jump to content


Photo
- - - - -

Buckaroo's Cloning Tek...{merged}


  • Please log in to reply
41 replies to this topic

#21 BuckarooBanzai

BuckarooBanzai

    The Abe Vigoda of Mycotopia

  • Moderator
  • 8,968 posts

Awards Bar:

Posted 07 January 2006 - 01:51 AM

DUDE...temper...I remember something about that from the wayback machine. Something about quenching and crystaline structures and hardness. Doesn't that also have something to do with the "rainbows" that you get in tempered metal that is heated red hot? Or is that hardened metals?

Ah, heck, I'm mixing ideas. I gotta go read some stuff about tempering and oxidation and hardness.

#22 suckerfree

suckerfree

    GATE KEEPER

  • OG VIP
  • 3,032 posts

Awards Bar:

Posted 07 January 2006 - 10:39 AM

an electrical current is used in the temper/coating process to get the rainbows.

#23 Guest_freakachino_*

Guest_freakachino_*
  • Guest

Posted 08 January 2006 - 07:23 PM

Hippie, I had to test it a few times for my microwave power, and I found a straight 5 minutes on high works like a charm every time. Different power levels on microwaves do make a difference so I just found the 5 minute mark to work well for me.

#24 BuckarooBanzai

BuckarooBanzai

    The Abe Vigoda of Mycotopia

  • Moderator
  • 8,968 posts

Awards Bar:

Posted 09 January 2006 - 11:17 AM

So, basically, you clean the microwave, put in your lightly capped LC containers and run it on high for 5 minutes?

Suckerfree - could you elaborate on the electrical/rainbow process? Or just give me a technique or link...

#25 Guest_freakachino_*

Guest_freakachino_*
  • Guest

Posted 09 January 2006 - 04:50 PM

Basically, I clean the microwave. Get my jar(s) cleaned and ready with the karo/water and marble. I like to really stir it up so the karo gets mixed well with the water. I use plastic lids for these so I can microwave. Lay the cap on top, then stick in and nuke for five minutes. After a few minutes of cool down, I slowly open the door, and tighten the lid down. Sometimes I've tightened the lid too much and when I'm ready to drop the clone tissue in I've had trouble getting the lid open to drop it, so now I make sure its tight enough but not so much I can't open it to add the tissue. The plastic lids have been a great addition to my mycology hobby because they are one piece and microwaveable. I just like cloning in lc's like your tek, easier for me and quickly gives me a lot of clone mycelium in 2-3 weeks time. I really think Hippies Microwave tek for lc's is best for me. Pressure Cooking my lc jars never seemed to work for me. Hippies tek has always worked for me, but then all of Hippies teks have worked for me when I've tried them! Rock On Hippie!

Do you like half pints or pints for your liquid culture clone jars? I've only used half pints but am thinking pints might work a little better and give more mycelium. Might add a bit more time though.........

#26 BuckarooBanzai

BuckarooBanzai

    The Abe Vigoda of Mycotopia

  • Moderator
  • 8,968 posts

Awards Bar:

Posted 09 January 2006 - 08:02 PM

I am going to try the microwave thing. Firing up the pressure cooker just to do two or three clone jars is annoying (and wasteful).

I started with half pints, but I only use pints now. I fill the jar about 1/2 full so I have plenty of air for growth without needing any kind of venting. How fast your LC will colonize seems to be more a factor of how much tissue you start with and how often you swirl the jar than size. The pint vs. 1/2 pint question seems more a matter of how much LC you want to end up with (his eyes glaze over as he imagines using 1/2 gallon jars for LCs).

10mL of thick clone water swirled 3-5 times a day will colonize a pint in a week or so. Bluehelix has show that with a mechanical stirrer, that time frame can be shortened to 2-3 days.

About 5-6 days in, when the jar is %50-%60 colonized, I shake the living CRAP out of it. This aerates the water and breaks up the mycelium for better "syringe suck up" later on.

Once I have a good container of clone water in distilled water, I use a small amount of it to start LCs and toss the clone original in the refrigerator. The LC starts the next generation and I maintain a "young" master culture to work from.

How do you handle long term culture storage? What's the oldest culture you have successfully worked from?

#27 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 15 February 2006 - 02:15 PM

OK. I've read a good deal of the archives on cloning TEK's (first-timer). Please direct me to the most widely used and beloved nowadays.
I am a rather simple person with no agar in the cupboards, what would you recommend? Karo? Marbles? Blending? Praying? Cap cuttings? Stem cuttings?
I'm utterly confused :special:

#28 Guest_dial8_*

Guest_dial8_*
  • Guest

Posted 15 February 2006 - 02:41 PM

I like using karo and water at a mix of 1 teaspoon karo to 100ml of water. I pc mine at 15 psi for 20 minutes some use a microwave on high for about 3-5 minutes with great success.

After it has cooled and you have selected the fruit you want to clone you need to do the rest of the work in a glove box. With fruit, karo jar, small dish for hydrogen peroxide, scapel, and tweezers inside the clean glove box you can begin work. With latex gloves on I like to pull the base of the stem in half long ways to reveal the inner tissue. I then take my scapel and cut a small portion of the inner tissue near the base of the stem. Then with tweezers I take the tissue drop it in the dish with hydrogen peroxide for 5 seconds and then transfer to the karo jar (no need to rinse the h2o2 off). I usually make at least 3 jars just to be on the safe side.

#29 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 15 February 2006 - 02:45 PM

Excellent. This is what I thought may be best for me but I just wasn't sure after reading so much info. A person just gets frazzled after a while.
Thanks so much for your help.:cool:

#30 Guest_dial8_*

Guest_dial8_*
  • Guest

Posted 15 February 2006 - 02:55 PM

No problem, man.
Here is a link that may help. [broken link]

I do not cut and peel like nan describes in this link. Like I said, I pull the stem open along the mushroom's long axis to reveal the inner tissue. Not only is this approach cleaner, imo, but it also leaves the inner tissue frazzled so more mycelium will come into contact with the karo water.


Edited by Sidestreet, 02 October 2016 - 03:44 PM.
removed broken link


#31 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 15 February 2006 - 03:07 PM

While you're on a roll......
can you then take some of this solution after its' incubation period (how many days do you let it sit?) and inject a syringe-full into another Karo jar or do you always start with a fresh piece of innards for each jar of solution?
Again, thank you-thank you:hippie:

#32 Guest_dial8_*

Guest_dial8_*
  • Guest

Posted 15 February 2006 - 03:16 PM

Innitially the 3 to 4 jars I do are all done with tissue, but if you are wanting to expand your mycelial mass into more lc's then yes you can take some from your original karo jar/jars and transfer via syringe to more karo jars.

I usually let mine incubate for 14 to 20 days. If at that time I am not ready t use it i stick it in the fridge. They can stay good in the fridge for quite a while.

#33 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 15 February 2006 - 03:42 PM

Does a person ever get "bored" cloning the same strain, same "big" shroom over and over again? Same flushes, same size, same same same? Do you ever revert to multi-spore just to see what you may come up with as a new host? Just curious... my mind usually can't stay focused in just one direction for any length of time, so I think I'll find myself enjoying this cloning thing for a while but will end up back from where I first started, if for nothing else but new visuals in the chamber:cool:

#34 Guest_dial8_*

Guest_dial8_*
  • Guest

Posted 15 February 2006 - 03:50 PM

Multispore is very important as kind of a reset button. Too much cloning can cause many problems. I do not clone the fruit produced by a cloned fruit. And expanding the mycelial mass especially cloned mycelium to far away from the original sample can sause degeneration. So yes, it is very important to still do ms inoculations.

#35 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 15 February 2006 - 04:02 PM

:) Awesome. Now I fully understand how to go through the process. I'm really looking forward to a bit of experimentation of my very own! Thank you again.

#36 python

python

    Former Member

  • Banned Member
  • 2,655 posts

Posted 15 February 2006 - 09:36 PM

flowhood/glovebox, interior tissue, agar/karo................cloning

#37 igetit

igetit

    Mycophage

  • Expired Member
  • 150 posts

Posted 16 February 2006 - 01:21 AM

im not sure if hippie posted a link to his hydrogen peroxide cloning, but I tried it and it is totally sucessful.

Inner tissue blended in a hydrogen peroxide / water solution -----> innoculation of sterilized jars -------> easiest way to clone that I've found.

#38 pcubie58

pcubie58

    Mycophiliac

  • Expired Member
  • 24 posts

Posted 16 February 2006 - 03:37 AM

This has all been very interesting reading, especially Python's link on the reproductive cycle. Just can't get enough of that!
I have 26 casings in fruiting chambers at the moment and so came the question about cloning for my next project...I'm wanting to try the Straw Log.
I've always multi-spored it and so it's time to do something different. I can't wait to see which strain comes up first and how aggressively.
I may try both the Karo and Cardboard teks for comparison. Either one seems easy enough!
I appreciate all your inputs. It's great to come home from work and have your hobby questions answered:hippie:

#39 Hippie3

Hippie3

    DUNG DEALER

  • Founders
  • 40,642 posts

Posted 18 February 2006 - 09:35 AM

long term LC store i use the fridge so metabolism slows.
i've heard of cultures 2 years old being revived
but they grew out with diminished vigor.

archive material

#40 viraljimmy

viraljimmy

    Mycotopiate

  • Expired Member
  • 452 posts

Posted 18 February 2006 - 01:56 PM

Has anybody tried dehydrating cultures for
long-term storage?

I was thinking with a little liquid or substrate,
allowed to dry in a jar with a poly or tyvek filter,
that it might last indefinitely.

Then it could be rehydrated and used to
innoculate. Would drying the culture damage
its future potential?




Like Mycotopia? Become a member today!