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Offlinezamora333
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Tissue sample
    #16169162 - 05/02/12 12:48 AM (12 years, 8 months ago)

So I don't know if anyone has tried this before but I just did and had some awesome success with it. It's way easier than going through that hassle of cutting inside for a tissue sample. I'm going to write up a step by step pictorial once I get another fruit that I like and show you guys. I cook a lot and I kind of got this idea of trying this when I was cutting some meat.

Pretty much you just grab a nice fruit that looks juicy and sweet  :tongue2:

Sterilize your work area as well as a half pint jar

Grab a nice sharp blade and put everything in a glove box or your work area and make sure everything is clean.

Now assuming that you have a decent size fruit you want to cut off the cap first. I never have contamination issues because I pick the fruit and put it directly into the glove box. Cut that cap off your mushroom. Next you want to slice up the stem of the mushroom like you would a carrot, make sure your knife is super sharp otherwise you might have some problems when cutting. Once you have the stem cut into the desired amount of pieces put them in the jar. Now take your cap and cut that into a grid an put them into the jar as well.

The tissues will be good for at MAX three days. I recommend using them immediately.

Now if you're me and you got a nice fruit you should have roughly around 30 pieces for 30 jars. One piece for each jar of grain  :cookiemonster:

Works like a charm, and it's an easier way of dealing with tissue samples.

As I said step by step pictorial tek coming soon from start to grain completion. :thatsinteresting:

Let me know what you guys think!


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All said and done are not my words, all pictures are fictional, all questions are rhetorical. In fact this is an optical illusion.

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InvisibleBeefy1
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Re: Tissue sample [Re: zamora333]
    #16169174 - 05/02/12 12:53 AM (12 years, 8 months ago)

You saying you just cut up a mushroom and use that like agar wedges to inoculate a grain jar?


sounds like a recipe for contams.

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OfflineThe Influence
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Re: Tissue sample [Re: zamora333]
    #16169177 - 05/02/12 12:54 AM (12 years, 8 months ago)

Quote:

zamora333 said:
So I don't know if anyone has tried this before but I just did and had some awesome success with it. It's way easier than going through that hassle of cutting inside for a tissue sample. I'm going to write up a step by step pictorial once I get another fruit that I like and show you guys. I cook a lot and I kind of got this idea of trying this when I was cutting some meat.

Pretty much you just grab a nice fruit that looks juicy and sweet  :tongue2:

Sterilize your work area as well as a half pint jar

Grab a nice sharp blade and put everything in a glove box or your work area and make sure everything is clean.

Now assuming that you have a decent size fruit you want to cut off the cap first. I never have contamination issues because I pick the fruit and put it directly into the glove box. Cut that cap off your mushroom. Next you want to slice up the stem of the mushroom like you would a carrot, make sure your knife is super sharp otherwise you might have some problems when cutting. Once you have the stem cut into the desired amount of pieces put them in the jar. Now take your cap and cut that into a grid an put them into the jar as well.

The tissues will be good for at MAX three days. I recommend using them immediately.

Now if you're me and you got a nice fruit you should have roughly around 30 pieces for 30 jars. One piece for each jar of grain  :cookiemonster:

Works like a charm, and it's an easier way of dealing with tissue samples.

As I said step by step pictorial tek coming soon from start to grain completion. :thatsinteresting:

Let me know what you guys think!



Looking forward to the pictorial :awethumb: I was thinking of doing something similar but transferring to an LC as I don't have a PC right now to work with grains.

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InvisibleExiledmonkies
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Re: Tissue sample [Re: zamora333]
    #16169181 - 05/02/12 12:56 AM (12 years, 8 months ago)

Sounds simple enough but only problem I see is the time limit and higher risk of contam. If it works well for you, one up to you! I prefer to inoculate a malt agar medium with a tissue sample and use that for the rye. That way you know from that start if there is contam rather then throwing something contaminated into a rye jar and wasting a extra 1-2weeks. :smile:

:mushroom2:

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Offlinezamora333
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Re: Tissue sample [Re: Exiledmonkies]
    #16169188 - 05/02/12 12:59 AM (12 years, 8 months ago)

I have a segregated room so I never have issues with contamination :wink:

And yes just like agar wedges except in grain jars. The thing that I like about it the most is it's very aggressive in the jars.


--------------------
All said and done are not my words, all pictures are fictional, all questions are rhetorical. In fact this is an optical illusion.

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Offlinezamora333
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Re: Tissue sample [Re: zamora333]
    #16169204 - 05/02/12 01:06 AM (12 years, 8 months ago)

Yeah Agar is boss but, I just started doing this as a way as a short cut from agar. I just found it unnecessary although I do really enjoy the science behind doing it :smirk:. I usually keep one grain jar as a master and once I run out I just transfer and make twenty more. I really think this will help beginners out a lot who either don't understand the concept of agar or can't afford to do it. Me I went all out I sunk about 1500 in this hobby thus far in supplies
:blowmybrainsout:

It was worth it though because I really enjoy it.


--------------------
All said and done are not my words, all pictures are fictional, all questions are rhetorical. In fact this is an optical illusion.

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OfflineThe Influence
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Re: Tissue sample [Re: Exiledmonkies]
    #16169215 - 05/02/12 01:10 AM (12 years, 8 months ago)

Quote:

Exiledmonkies said:
Sounds simple enough but only problem I see is the time limit and higher risk of contam. If it works well for you, one up to you! I prefer to inoculate a malt agar medium with a tissue sample and use that for the rye. That way you know from that start if there is contam rather then throwing something contaminated into a rye jar and wasting a extra 1-2weeks. :smile:

:mushroom2:



You make good points, but for someone like me that does not have the materials to work with agar or grains at the moment this really is the only viable option to make a clone/isolate. I will probably do it a little different but the idea is still the same.

I do all my work in a GB as well and I will cut the cap off of the fruit so as to not risk spores getting mixed in while I work. I will wipe the stem down with ISO and instead of just cutting the fruit up I will take a section out of the center of the stem that has not been exposed to open air and then introduce immediately to an LC.

Really I don't see how it's much more risky than making a syringe from a print :shrug:

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InvisibleOnePerEyeM8
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Re: Tissue sample [Re: The Influence]
    #16169378 - 05/02/12 02:08 AM (12 years, 8 months ago)

Looks like a nifty alternative when agar isn't an option! :smile:

Quote:

I do all my work in a GB as well and I will cut the cap off of the fruit so as to not risk spores getting mixed in while I work. I will wipe the stem down with ISO and instead of just cutting the fruit up I will take a section out of the center of the stem that has not been exposed to open air and then introduce immediately to an LC.




GB here too. I wish I had the spare dinero for a flow hood haha. Shit, I actually just remembered that I had a dream about getting a flow hood a couple nights ago.

I made some BRAFAS (just came across the tek, figured I'd try it out) the other day to clone some edibles. Pulled apart the stem with iso soaked, gloved hands to get at the inner tissue, then used a flame-sterilized pair of tweezers to take the tissue sample.

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InvisibleExiledmonkies
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Re: Tissue sample [Re: OnePerEyeM8]
    #16169393 - 05/02/12 02:15 AM (12 years, 8 months ago)

Quote:

OnePerEyeM8 said:
Looks like a nifty alternative when agar isn't an option! :smile:

Quote:

I do all my work in a GB as well and I will cut the cap off of the fruit so as to not risk spores getting mixed in while I work. I will wipe the stem down with ISO and instead of just cutting the fruit up I will take a section out of the center of the stem that has not been exposed to open air and then introduce immediately to an LC.




GB here too. I wish I had the spare dinero for a flow hood haha. Shit, I actually just remembered that I had a dream about getting a flow hood a couple nights ago.

I made some BRAFAS (just came across the tek, figured I'd try it out) the other day to clone some edibles. Pulled apart the stem with iso soaked, gloved hands to get at the inner tissue, then used a flame-sterilized pair of tweezers to take the tissue sample.


Build one yourself. You can get the tek from many websites online. Mine only cost me 250ish..

....If you already have a nice amount of decent power tools. :p

Edited by Exiledmonkies (05/02/12 02:21 AM)

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OfflineThe Influence
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Re: Tissue sample [Re: OnePerEyeM8]
    #16169402 - 05/02/12 02:20 AM (12 years, 8 months ago)

Quote:

OnePerEyeM8 said:
Looks like a nifty alternative when agar isn't an option! :smile:

Quote:

I do all my work in a GB as well and I will cut the cap off of the fruit so as to not risk spores getting mixed in while I work. I will wipe the stem down with ISO and instead of just cutting the fruit up I will take a section out of the center of the stem that has not been exposed to open air and then introduce immediately to an LC.




GB here too. I wish I had the spare dinero for a flow hood haha. Shit, I actually just remembered that I had a dream about getting a flow hood a couple nights ago.

I made some BRAFAS (just came across the tek, figured I'd try it out) the other day to clone some edibles. Pulled apart the stem with iso soaked, gloved hands to get at the inner tissue, then used a flame-sterilized pair of tweezers to take the tissue sample.



Dude that is fucking awesome :thumbup: Thanks for showing me that man +5! (well and updated rating as you already are the one that taught me rice bran is superior to brf some months ago)

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Offlinezamora333
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Re: Tissue sample [Re: The Influence]
    #16169468 - 05/02/12 02:55 AM (12 years, 8 months ago)

Nice! Gonna give this a try as well.


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InvisibleExiledmonkies
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Re: Tissue sample [Re: zamora333]
    #16169513 - 05/02/12 03:19 AM (12 years, 8 months ago)

I'm with those two. I've never tried that tek before. Def trying it now!:cool:

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InvisibleDoc_T
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Re: Tissue sample [Re: Exiledmonkies]
    #16169848 - 05/02/12 07:02 AM (12 years, 8 months ago)

zamora, I'm glad this works for you, but in general it's bad advice.
Newbs in particular are going to have trouble with contams. Even you will sooner or later, though it's working now.
Not all contaminants are visible or grow rapidly, you could have a master jar that looks ok but the final tubs never fruit.
Dropping the slice onto an agar plate, then transferring a wedge to your grain jar, gives you one more assurance of a clean culture.

Beats hell out of the needle-and-LC route though, I'll give you that. :thumbup:


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You make it all possible. Doesn't it feel good?

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Offlinerohr
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Re: Tissue sample [Re: Doc_T]
    #16171275 - 05/02/12 01:18 PM (12 years, 8 months ago)

Very interesting!

So the fruits produced from a tissue sample transfer to grain have similar characteristics to the fruit that tissue was taken from, just like an actual clone on agar? Fungus never ceases to amaze me. I'm so going to try this as soon as I have fresh fruits from the grain jars I inoculated just the other day, cannot wait.


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Re: Tissue sample [Re: rohr]
    #16171397 - 05/02/12 01:51 PM (12 years, 8 months ago)

A tissue clone is the same whether you grow it out on agar or grain or brf or in an LC.
It's not usually a genetic isolate though. Typically a mushroom will contain genetics from multiple individual organisms.
These will sector out on agar and can be isolated if desired. (Same idea as starting from spores).
It's not really like a coral colony but that's a useful way to think about it, smaller creatures making a larger whole.


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Offlinedirtdogg
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Re: Tissue sample [Re: Doc_T]
    #16171469 - 05/02/12 02:09 PM (12 years, 8 months ago)

you ever check out this?

9er cloning tek



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the quieter you are the more you can hear

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InvisibleDoc_T
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Re: Tissue sample [Re: dirtdogg]
    #16171795 - 05/02/12 03:16 PM (12 years, 8 months ago)

That wasn't a very good tek when it was posted ten years ago, and it hasn't aged well at all.
Requires the same sterile technique as agar work, without any of the benefits of agar work.


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Offlinezamora333
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Re: Tissue sample [Re: Doc_T]
    #16172535 - 05/02/12 05:56 PM (12 years, 8 months ago)

Yeah agar is where it's at, I just kind of developed this as a substitute... Pics and video coming soon!


--------------------
All said and done are not my words, all pictures are fictional, all questions are rhetorical. In fact this is an optical illusion.

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Offlinedirtdogg
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Re: Tissue sample [Re: zamora333]
    #16172651 - 05/02/12 06:18 PM (12 years, 8 months ago)

Quote:

zamora333 said:
Yeah agar is where it's at, I just kind of developed this as a substitute... Pics and video coming soon!




to each their own. you are correct sterile is the way to eat and breathe in this hobby, and ive never used agar yet. i just know what works for me.

1 ml of this would colonize faster the 1 ml of a normal LC is all im really saying. no tissue sample from multi spore LC, clone, or syringe would give you a isolate unless you did agar work. so yes you are correct just wanted to throw out another option for them.

http://www.shroomery.org/forums/showflat.php/Number/11353633#11353633

this dude seems to do well from what i've read with 9er cloning tek and other various methods.


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the quieter you are the more you can hear

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Offlinezamora333
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Re: Tissue sample [Re: dirtdogg]
    #16172685 - 05/02/12 06:23 PM (12 years, 8 months ago)

Yeah I actually looked over that before I like the hepa box he made. I also like the blending idea, was thinking about knocking up some BRFs and trying it out the only thing to me is that it seems it would take the same amount of time to just knock up a Rye jar unless the growing performance is better than it'd be worth the wait for sure.


--------------------
All said and done are not my words, all pictures are fictional, all questions are rhetorical. In fact this is an optical illusion.

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