Pan Cyan Tissue Cloning....How?

[pvacant]

Anyone have experience getting Panaleous cyanescens tissue for cloning?
Just where DO you get spore free tissue from such small fruits?
Is the 9er tek (cloning) a possibility? Or is MS to agar, then isolating, the only option; cause i would rather clone a selected fruits tissue.....but HOW!?
thanx everyone..........

[joystik]

i cut a very small piece of stem and soaked it in peroxide for a couple minutes, then to agar.

[Shroomette]

How I do my cloning is first take and make sure the cleanroom is very sterile by spraying oust/lysol in there and let that sit in there for 20 mins. Then, spray some more while in there, wash my hands with 91% ISO, and take the mushroom that your cloning and wipe it down with ISO. then I sterilize my knife or scissors that I plan on using. Cut a slit up the to the middle of the stem, and using a pair of sterlized tweezers and peel out the tissue from stem close to the cap. Then, from there drop it into a LC. I used the Honey/karo LC by irishlion.

Good Luck pvacant, here is some good vibes for cloning.

[pvacant]

thanx you two.

now joystick, are you saying you cut a piece of stem, and drop the whole thing in the H2O2, the outside of the stem as well? and if so does that usually involve subsequent dish transfers to clean it up?

and Ette, i myself haven't had much luck getting tissue to take in an LC, spores are fine but not tissue. It always stalls and just disintegrates into fine particles that collect on the bottom. but i looked up Irishlion's LC tek and i do like it. I'll give it a whorl too, thanx for all the advice. i just can't seem to clone a fruit as small as pans....still way to clumsy with agar work for that tiny of a stem! hell even trying to get a clean sample for the LC is gonna be tough. oh well i'll try it anyway.
have a good weekend

[whatchamacallit]

The peroxide should help keep it clean if some of the outside of the stem is used. It's best to try and get it from the inside of the stem, but not always possible. SWIM would suggest taking several clones on several different plates for a better chance of success. The iso wipe and peroxide soak should help a good bit with getting a clean sample transferred. SWIM also likes using agar to clone, and finds it easier than using an LC, many times, altho trying both wouldn't hurt. After all, the more teks tried, the more chances one has for success. The agar wedges can be thrown in a jar, to inoculate, and these jars used for g2g's to multipy the spawn. SWIM finds this as his preferred method.

[joystik]

pvacant: Yes. Usually copelandia stems are too thin to split and take inner tissue, so I used the method I described above using the whole piece.

What I do now when I dont want to get into agar making is, I take a small jar, like a baby food jar and cook 1 or 2 spoonfulls of millet or popcorn. PC it and let it cool. Once that is ready, i cut the stem, leave it in peroxide for 1 minute and then throw it in the jar and close it. I like to do a few of them just in case. After a few days the mycelium will start to jump onto the grain. When u see a few clean fully colonized grain kernels, using a sterile loop or small spoon, take 1 (one) clean kernel very carefully and you can put that in agar or, like I do, use that to inoculate a Sterilized LC. Then u have your LC from cloned tissue.

[Jose_sixpac]

Mucho's grassy, farie Goddess...

Swim is gonna do this manana...


Just one question...swim has wild pannies that are prett small, still one fresh stem biopsy per LC?



swim is pissin their pannies in anticipation.


swim is still gettin hippsters fire and forget kit for the invetro project swim has in mind for some B+ters, when I am a little more aged here. Or can I order Be-4 having "T" status?

-j-