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Offlineink
heffalump
Registered: 03/24/04
Posts: 21
Last seen: 11 years, 11 months
Cardboard cloning tek?
    #2817961 - 06/22/04 06:51 PM (12 years, 8 months ago)

Can anyone point me in the direction of a cardboard cloning tek? thanks


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Offlinecharvo
JOURNEYMAN
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Registered: 03/09/03
Posts: 654
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Re: Cardboard cloning tek? [Re: ink]
    #2818791 - 06/22/04 10:39 PM (12 years, 8 months ago)

I havn't seen a tek on cardbroad cloning here, but i can tell how.

first cut a 1inchx1inch piece of cardbroad, then tear the side off exposing the ribbed section underneath.
place the cardbroad in a sealable container and fill with a 1/4 inch of water.microwave for about 3 minutes.pour out any access water that remains you don't want the cardbroad to wet.just needs to be moist.place the wedge of your choice on the ribbed section of the cardbroad.thats it man you should see grow in a few days.

see some example here.the update should have a nice distribution of cardbroad cloning with pictures.

http://www.shroomery.org/forums/showflat...;o=&fpart=1


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Offlineink
heffalump
Registered: 03/24/04
Posts: 21
Last seen: 11 years, 11 months
Re: Cardboard cloning tek? [Re: ink]
    #2818886 - 06/22/04 11:03 PM (12 years, 8 months ago)

Thanks


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OfflineGr0wer
always improving
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Registered: 09/16/03
Posts: 5,807
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Re: Cardboard cloning tek? [Re: ink]
    #2819378 - 06/23/04 12:52 AM (12 years, 8 months ago)

But thats not sterile is it?


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Offlineink
heffalump
Registered: 03/24/04
Posts: 21
Last seen: 11 years, 11 months
Re: Cardboard cloning tek? [Re: Gr0wer]
    #2819421 - 06/23/04 01:07 AM (12 years, 8 months ago)

guess the microwave kills all the bad stuff, could you use H202 to dip the wedge in?


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OfflineGrouch
el brujo

Registered: 03/05/03
Posts: 116
Loc: New Mex
Last seen: 10 years, 9 months
Re: Cardboard cloning tek? [Re: ink]
    #2819906 - 06/23/04 04:26 AM (12 years, 8 months ago)

question, how long can this cardboard clone be viable, can it just be stored inside a jar in the fridge (im assuming yes)? and do you just cut a cardboard wedge out and do a grain transfer or would agar be recommended?

thanks


--------------------
The Mexican Wizard


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InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Cardboard cloning tek? [Re: Grouch] * 1
    #2821770 - 06/23/04 06:55 PM (12 years, 7 months ago)

Quote:

Grouch said:
question, how long can this cardboard clone be viable, can it just be stored inside a jar in the fridge (im assuming yes)? and do you just cut a cardboard wedge out and do a grain transfer or would agar be recommended?




The mycelium on the cardboard should last as long under refrigeration as mycelium produced with other methods (months, years, decades - depends on the storage technique). It should be well sealed, as it can easily dry out. Yes, you can cut a wedge and go directly to grain, or transfer it to agar first. Cardboard works fine for cloning, although not all species will take to it. It is especially useful when you have a single large mushroom but do not want to bother cooking up a batch of agar. This is what I do, and is much faster.

Quote:

Gr0wer said:
But thats not sterile is it?



It is sterile if you follow the proper precautions. Sterility is up to you...


Here is the method as outlined in my thread that charvo linked to:


1. - First separate one of the smooth sides, exposing the internal ribbed surface. Notice it does not peel off completely, leaving fuzzy remnants of cardboard.
2. - Clean off all remnants of the flat piece, making a smooth surface for maximum contact area. Plus, you don?t want left-over fibers that take forever to colonize sticking up and drying out.
3. ? I take a half pint jar using a plastic lid with a poly-fill wad and fill ? full with water. Insert the prepared cardboard and microwave. I microwave on full power until it boils, then turn the temp down so it boils, then stops, and continues this cycle for 10-15 minutes. Allow to cool in the microwave, then place an alcohol soaked coffee filter over the lid, secured with a rubber band. This will protect it during transport back to the lab. Microwaving in solution not only sterilizes the mix and accelerates hydration; it also cooks out almost all of the undesired cardboard glue whereas soaking alone does not. It is also much faster than pressure cooking, which is really not necessary given the selectivity of cardboard.
4. - Now it is time to slice into the mushroom. There are 2 ways to go, either a piece of the stem, or flesh from the cap. Rip the cap in half, exposing the thick flesh above the gills then cut out a small piece with a sterilized scalpel. Make sure you do not get any of the cap surface or any of the gill tissue. These places can usually harbor contaminant spores and bacteria.
5. - Here the extracted tissue is transferred to the cardboard in a petri dish. The mushroom tissue should be pushed into the recess of the rib forcefully, maximizing contact area. It is then sealed and incubated.

It also should be noted that after transport back to the lab, the container is again wiped down with isopropyl alcohol when in the flowhood airstream to again insure no contaminants will make it into the culture. Like I said, the cardboard is very selective and assuming you microwave it long enough, it should be perfectly sterile. All the steps you take after it is cooked are the ones that will introduce contaminants. It is up to you to insure you make this process as clean as possible whether you have a glovebox, flowhood, etc. I cannot recommend anything open air - As I said, sterility is up to you. If you want to pressure cook the cardboard, fine. But IME it is not necessary given the selectivity of the cardboard (and if I were to PC something, I might as well make agar).

I have had nothing but 100% success with cubensis like this, with a 0% contamination rate after transferring to agar or grain (I have used cardboard many more times than I could ever keep track of, but I know every attempt was successful). I actually prefer this method over agar for cubensis tissue cloning. At least for producing viable tissue for future agar work. This technique has also worked saving a blue oyster culture from black pin mold also. It is also very important to not over-saturate the cardboard...

Give it a try, WTF have you got to lose? An hour of time, a 1"x1" square of cardboard, and a tiny chunk of a mushroom...


--------------------
To give is to live...



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Offlinecharvo
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Registered: 03/09/03
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Re: Cardboard cloning tek? [Re: ATWAR]
    #2821891 - 06/23/04 07:55 PM (12 years, 7 months ago)

this tek should be archived.IMO

I don't recall see one in FAQ lately at the shroomery, althought i don't read the FAQ's much.  i do remember reading one long time ago somewhere.i guess it could have been at the shroomery, but i don't remember.


lets send out big thanks to ATWAR for taking the time to photograph and document this tek. :thumbup:


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Offlinedoc34
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Registered: 02/14/04
Posts: 2,660
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Re: Cardboard cloning tek? [Re: charvo]
    #2821906 - 06/23/04 08:02 PM (12 years, 7 months ago)

Quote:

  this tek should be archived.IMO







I agree!!

Great Job ATWAR  !!!!

:mushroom2: :mushroom2: :mushroom2: :mushroom2: :mushroom2:

:cool: :thumbup:


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


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InvisibleMykey
spectraltraveler

Registered: 04/07/04
Posts: 542
Re: Cardboard cloning tek? [Re: doc34]
    #2822063 - 06/23/04 08:59 PM (12 years, 7 months ago)

I couldnt agree with you guys more! As always,damn good job ATWAR!!

MYKEY


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InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Cardboard cloning tek? [Re: Mykey]
    #2822170 - 06/23/04 09:39 PM (12 years, 7 months ago)

I know its been done before...
I just documented it quick for the log...


--------------------
To give is to live...



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OfflineGrouch
el brujo

Registered: 03/05/03
Posts: 116
Loc: New Mex
Last seen: 10 years, 9 months
Re: Cardboard cloning tek? [Re: ATWAR]
    #2822880 - 06/24/04 02:54 AM (12 years, 7 months ago)

Thanks for taking the time for the tek and pics, will be put to good use!


--------------------
The Mexican Wizard


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OfflineJonno
NewPin

Registered: 06/13/03
Posts: 15
Last seen: 12 years, 4 months
Re: Cardboard cloning tek? [Re: ATWAR]
    #2833864 - 06/27/04 05:23 PM (12 years, 7 months ago)

What a great idea! I've been trying variations of this tek out and have found that the cardboard doesn't need boiling, just dampen with 10:1 water/peroxide(3%)
I did have concerns that bacteria would be unable to colonize the card but infect my grain, this hasn't happened but I use popcorn as spawn, and my jars are fully colonized in a week. I guess the bugs don't have a chance.
A wet sandwich box in a mass of polythene bags and trash got through a recent customs inspection after a trip to Amsterdam. The small sections I had dotted around it are doing fine. The customs guy was more interested in the stale coffee in my Thermos.


--------------------
Use Popcorn Your shrooms will love you back!


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InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Cardboard cloning tek? [Re: Jonno] * 1
    #2834668 - 06/27/04 10:46 PM (12 years, 7 months ago)

If you read the most important part about boiling it is to reduce the glue content in the corrugated cardboard. Sterilization is up to you...


--------------------
To give is to live...



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Offlineadizzle
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Registered: 01/01/11
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Re: Cardboard cloning tek? [Re: charvo]
    #13989847 - 02/19/11 02:39 PM (6 years, 5 days ago)

Pretty excited about trying this today


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Offlinemycocozm


Registered: 05/13/10
Posts: 181
Last seen: 1 year, 8 months
Re: Cardboard cloning tek? [Re: adizzle]
    #14011182 - 02/23/11 02:23 AM (6 years, 2 days ago)

Why isn't this more commonplace if cardboard favors mushroom mycelium over mold and bacteria?  Is the colonization rate much slower?


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Offlinemycocozm


Registered: 05/13/10
Posts: 181
Last seen: 1 year, 8 months
Re: Cardboard cloning tek? [Re: mycocozm]
    #14011232 - 02/23/11 02:30 AM (6 years, 2 days ago)

While we're at it, what about germinating spores this way? I'd way rather see cardboard get reused than keep buying seaweed.


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Offlineadizzle
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Re: Cardboard cloning tek? [Re: mycocozm]
    #14016190 - 02/23/11 10:28 PM (6 years, 1 day ago)

It seems to be working so far, with noticeable changes within 3-4 days...


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Offlineweaksause
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Registered: 08/26/14
Posts: 254
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Re: Cardboard cloning tek? [Re: mycocozm]
    #20611038 - 09/24/14 01:38 PM (2 years, 4 months ago)

Quote:

mycocozm said:
While we're at it, what about germinating spores this way? I'd way rather see cardboard get reused than keep buying seaweed.




good question ! is there an answer ?
would be really sweet :syringe: :rocket:


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OfflineKizzle
Misanthrope

Registered: 08/30/11
Posts: 8,647
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Re: Cardboard cloning tek? [Re: weaksause]
    #20621170 - 09/26/14 01:38 PM (2 years, 4 months ago)

I don't think they germinate on plain cardboard. Never know until you try though:shrug:

If you're looking for an alternative to agar I know you can germinate spores on BRF+water.


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


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