50 replies to this topic

[dial8]

Well I have only cloned cubes and I did use inner stem tissue. I just though it may be easier to get inner tissue from the cap since it is a bit thicker than the stem.

Yeah, I would not try to clone a section of the gills that was loaded with spores. That would be basically the same thing as a multi spore grow.

[golly]

Off to a bang..!..How long did they take to show...?

[tbonus]

good luck fella's

[Elf Salvation]

cool!!!!!!!!!:space:

[spacecake]

Off to a bang..!..How long did they take to show...?

After two days I saw sign of new growth on the stems Golly !

[golly]

Great..I'm guessing mine take longer because i use the 35% peroxide...
may the force be with u...heh....

[mycot]

Inspired by Golly's thread "Pan Cyan clone experiment" I've been experimenting with cloning wild Pans recently.

As anybody who has tried, getting inner tissue from Pans, cap or stem it's a bitch not that it cant be done.

I therefore worked with stem sections splitting them down the center thinking to expose more inner tissue. This was dunked in 3% h2o2 for one minute and transfered to small baby food jars of sterilised millet.
I chose millet because it could be quickly prepared. It also provides many points of contact with the cloning tissue. I could have chosen WBS but having a single grain would be more specific.You'll see why I mention this in a minute.

Fourteen jars and two pan species were involved.
Additionally another six jars of an agar substitute comprised of BRF and water were prepared.
The result for all 14 jars was healthy myc growth on the grain with no signs of contamination on same.
The 6 jars on the other hand were a total loss. All sucumbed to bacterial contamination.

What I surmise from this at this point is that 3% h202 is insufficient to totally sterilise the tissue. The reason that the 14 grain jars did not contaminate was because of less points of contact with the tissue and because of more speceficity.

A small amount of collonised grain away from the transfered stem section was innoculated onto my favorite Pan spawn (Popcorn).
All looks good so this isn't a bad way of working. :D

[golly]

Interesting,,,U sometimes see that same effect in normal sterilized grain jars, where most do fine but one thats a bit wetter or less ventilated will slightly favour bacterial growth...Keep us posted Mycot ,thanx...

[spacecake]

A small update...

Yesterday I saw myc. growth on different spots..don't now why ?
Hopefully it aren't spores germinating (can't be ?!)
Or myc network is growing inside out ,..so I can't see.

Whatever it is,..things seem to go wel !

Attached Thumbnails

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[eu_por_exemplo]

Pretty vision of the beginning of the settling. Pans are fantastic.


:eusa_clap

[golly]

Good progression there,,,, I've also noticed alternate germination sites in clone jars..At first i thought it must be mold but i guess microscopic fragments break off durin the initial jar shake...

[reverend trips]

I wouldn't rule out the possability of it being spores that have clung on to the stem, seems like a logical explanation as well, I hope not, but how could you know for sure?

[spacecake]

I'm glad I am not the only one with this strange alternate germination...

I'll keep posting !

[mycot]

Interesting,,,U sometimes see that same effect in normal sterilized grain jars, where most do fine but one thats a bit wetter or less ventilated will slightly favour bacterial growth...Keep us posted Mycot ,thanx...

Totally agree Golly. In using the technique described every effort should be made to ensure the clone jars are not overly wet and therefore favor bacterial growth. Better on the dry side.
With overly wet grain,at best one gets very slow mycelial growth, at worst loss of the jar to contam.

[Bobcat]

I'd note that the liquid pooling on the bottom has a whitish color. Could very well be starches off the grain. Also could be some bacteria. Maybe try some verm, coir or similar on bottom to collect excess liquid, as already suggested (added to bottom dry b4 sterilization). Its still very simple and yet effective.

Now, to continue this 'simplicity' idea.... Yur pretty advanced, Space! You ought to give agar a try before throwing it into the 'complicated' category! You have the skills and it isn't hard! Plus you wouldn't have to worry about those darn contaminated spores, if ya know what I mean!

[bluehelix]

Pan Cyans LOVE liquid cultures. They grow better in LC than most any other mycelium I know. Once they get in real substrates, the mycelium is a lot weaker which is strange. I am starting up some viets again and you know I am using LCs just like have worked for me many times in the past.

[spacecake]

Someday I will try Agar,but I haven't a clue how to get my hands on it,and it's not on my ''thirst thing to do'' list...

All went very good and the clones are now spawned/cased and in the incubator, I think I will see some shrooms in two days or so !

I have some Viets pinning at this moment too,no big flush but very,very healthy shrooms.. with the double layer casing !

Pictures soon !

[golly]

Glad it's going well for ya Space...What method did u use for transfering your clone myc to spawn jars...?

BH...I finally broke down and bought a magnetic stirrer for some edible cultures ...Was inspired by your last gro-log...Not ready to thankyou yet though as i havn't actually started...:lol:

[spacecake]

Glad it's going well for ya Space...What method did u use for transfering your clone myc to spawn jars...?

Methode ?..lol !, just quickly dumped the stem in the jar in open air on the kitchen table and shaked it very rough !

I heard Seven saying that his Florida strain doesn't do wel when cloning.. so I didn't bother to do things carefully !
And I find it more fun when it works this way !