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Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
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zzantium2999
Shroom Groomer



Registered: 03/22/11
Posts: 177
Loc: Toronto Canada
Last seen: 12 years, 2 months
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LC from live tissue
#14359194 - 04/27/11 01:13 AM (12 years, 9 months ago) |
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Can you make a liquid culture from live tissue out of the centre of the mushroom stem? I figured that way I can be sure to get a culture that only has one strain. Am I correct in assuming this, or is there an easier way to make sure that I'm only putting one strain in the liquid?
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A good friend would bail you out of jail, but your best friend would be the one sitting next to you saying, "damn that was awesome".
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BargainBab
Hey Dere Ho Dere



Registered: 04/04/11
Posts: 1,178
Loc: Callisto
Last seen: 10 years, 2 months
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You mean cloning? Just get some agar man
-------------------- Oh wow, GOOD Nyborg!
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Mushaboom
Teacher/Student



Registered: 06/21/10
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It works and there are teks for it.
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"What you think, you become."
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awakendone


Registered: 08/05/10
Posts: 824
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i've never tried it, but it's been done LC Cloning
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BargainBab
Hey Dere Ho Dere



Registered: 04/04/11
Posts: 1,178
Loc: Callisto
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Re: LC from live tissue [Re: awakendone]
#14359237 - 04/27/11 01:28 AM (12 years, 9 months ago) |
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Was looking for that link
-------------------- Oh wow, GOOD Nyborg!
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zzantium2999
Shroom Groomer



Registered: 03/22/11
Posts: 177
Loc: Toronto Canada
Last seen: 12 years, 2 months
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Re: LC from live tissue [Re: BargainBab]
#14359246 - 04/27/11 01:32 AM (12 years, 9 months ago) |
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did you even read the question? I don't want to save my culture on agar. I want to make liquid culture so I can inoculate with that. The question was...Will it work with live tissue. Argh sometimes it's like people are just posting, to get more posts under there belt, and don't even read the questions. Or there are those ass holes who just post "yea I agree."' Who the hell asked if you agree. they asked a question, not if you agree with the last post. Sorry for the rant, but it just pisses me off when I need something answered and I see posts that have nothing to do with the question or topic.
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A good friend would bail you out of jail, but your best friend would be the one sitting next to you saying, "damn that was awesome".
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ar1es
Psychonaut



Registered: 07/09/10
Posts: 776
Last seen: 1 year, 3 months
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use the search engine
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ar1es
Psychonaut



Registered: 07/09/10
Posts: 776
Last seen: 1 year, 3 months
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Re: LC from live tissue [Re: ar1es]
#14359396 - 04/27/11 02:42 AM (12 years, 9 months ago) |
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only way to isolate is with agar
a piece from the center of the mushie will make clones but it wont be an isolate it will still contain may different genetics
or maybe im wrong
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Omnicracker
Crusted Trolltivator


Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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Re: LC from live tissue [Re: ar1es]
#14359986 - 04/27/11 08:21 AM (12 years, 9 months ago) |
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it will work, but cloning, even when done right can be very dirty. agar will make sure you didnt get any contams. if you go to LC direct, it could get contamed and you'll never know. it will work though. just put 2 marbles and some shards of glass in the jar to help break things up when it gets growing.
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Trippy_Smurf
Sketchy Mother Fucker




Registered: 02/14/11
Posts: 2,349
Loc: Smurf Villiage
Last seen: 2 years, 4 months
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You'll get attitude anytime you ask a question that has been answered 100's of times and can easily be found with the search engine.
You can clone the stem, which will only have one set of genetics, but there's a good chance the LC will be dirty/contaminated since the tissue sample is not sterile. Agar is used to make sure you're working with a clean sample before you waste your time and resources.
If you're going to skip the agar, make sure to noc up a few pf jars to test it before you use it on lots of jars.
-------------------- SECURITY: READ THIS! Chef: Kids, what did I tell you about drugs? Kids: There's a time and a place for everything, and it's called college. How to be a good shroomie How to grow mushrooms A collection of good links (may be outdated) How things should look How to pass a drug test
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Quote:
zzantium2999 said: Or there are those ass holes who just post "yea I agree."' Who the hell asked if you agree. they asked a question, not if you agree with the last post.
Agreed.
-------------------- You make it all possible. Doesn't it feel good?
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Sillicybin
Registered: 02/14/05
Posts: 2,134
Loc:
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Re: LC from live tissue [Re: Doc_T]
#14360104 - 04/27/11 08:56 AM (12 years, 9 months ago) |
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Quote:
Doc_T said: Agreed.
Doc_T for president. 
Can you go from live tissue to LC? Yes. Should you? No. You will almost certainly get an LC with bacterial contamination. LC is a contamination magnet. It's awesome when it's sterile, and a huge waste of time when it's not.
A better way to do this might be to toss a good biopsied fruit chunk into a grain jar using a glovebox. With that, you will have better chance of success because even if it does have some contamination on board, the mycelium might be able to overcome it in grain where it certainly cannot in LC. Then you can use Doc_T's method of making GLC once you've got a solid grain jar.
All of the above is the slacker way of doing things. If you really want to clone the right way, procure and learn to use agar.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: LC from live tissue [Re: Sillicybin] 1
#14360109 - 04/27/11 08:57 AM (12 years, 9 months ago) |
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Quote:
Sillicybin said:
Can you go from live tissue to LC? Yes. Should you? No. You will almost certainly get an LC with bacterial contamination.
Agreed. I never did get it to work, although lots of people do.
-------------------- You make it all possible. Doesn't it feel good?
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: LC from live tissue [Re: Doc_T]
#14360113 - 04/27/11 08:59 AM (12 years, 9 months ago) |
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Also, a clone is not an isolate. Generally there will be several, possibly many, substrains in a tissue sample.
-------------------- You make it all possible. Doesn't it feel good?
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: LC from live tissue [Re: Doc_T]
#14360126 - 04/27/11 09:03 AM (12 years, 9 months ago) |
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I've dropped colonized popcorn grains, bits of colonized substrate, core tissue, pins, agar into liquid culture with success.
There's just a lot of failure that goes with it. So u make several and toss the ones that turn cloudy. Core tissue and agar can be used without a peroxide dip, but any other kind of tissue transfer to lc is going to need to be sterilized in h202. If you go over 20 seconds tho there is a tendency to kill the tissue.
The reason people are encouraging u to go to agar as an intermediate step is to increase your chances of success. They aren't trying to be dicks.
If you do go tissue -> sugar lc , then make several at one time cuz some will probably turn on u. Remember that even the slightest cloudiness means it's toast.
GL!
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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Sillicybin
Registered: 02/14/05
Posts: 2,134
Loc:
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Re: LC from live tissue [Re: anonjon]
#14360148 - 04/27/11 09:09 AM (12 years, 9 months ago) |
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anonjon - I'm pretty sure that post right there has to be the nicest-toned one you've ever submitted. I'm ... shocked.... Keep this up and we're going to wonder if you got kidnapped and replaced by someone else. 
All of the suggestions are spot-on, exactly my experience with it. The H2O2 dip is extremely helpful in trying to clean up a tissue sample for LC - that's the only way I was ever able to get it to work, and even then I still got over 50% failure rates.
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gornyhuy
A Myth Intrepidly Met



Registered: 08/04/09
Posts: 2,933
Loc: USA
Last seen: 2 years, 2 months
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Re: LC from live tissue [Re: Sillicybin]
#14360174 - 04/27/11 09:18 AM (12 years, 9 months ago) |
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To take it one step farther, if you want to have some fun, do it right, and get great results:
1)tissue -> agar 2)clean & isolate your clone on agar 3)agar wedge to one jar of grain 4)make a grain LC "GLC" from that jar 5)inject inject inject!!!
Now you are pretty much guaranteed to be 1)cloned 2)isolated and 3)clean as a whistle.
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Trippy_Smurf
Sketchy Mother Fucker




Registered: 02/14/11
Posts: 2,349
Loc: Smurf Villiage
Last seen: 2 years, 4 months
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Re: LC from live tissue [Re: Sillicybin] 1
#14360230 - 04/27/11 09:34 AM (12 years, 9 months ago) |
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Quote:
Sillicybin said:
Quote:
Doc_T said: Agreed.
Doc_T for president. 
I agree.
-------------------- SECURITY: READ THIS! Chef: Kids, what did I tell you about drugs? Kids: There's a time and a place for everything, and it's called college. How to be a good shroomie How to grow mushrooms A collection of good links (may be outdated) How things should look How to pass a drug test
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: LC from live tissue [Re: Sillicybin]
#14360973 - 04/27/11 12:57 PM (12 years, 9 months ago) |
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Quote:
Sillicybin said: anonjon - I'm pretty sure that post right there has to be the nicest-toned one you've ever submitted. I'm ... shocked.... Keep this up and we're going to wonder if you got kidnapped and replaced by someone else. 
It's all my new gf's fault. She's makin me soft!
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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k00laid
NEMO


Registered: 05/03/10
Posts: 19,636
Last seen: 6 months, 5 days
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agar is for more than saving your cultures, OP..
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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Re: LC from live tissue [Re: k00laid] 1
#14361011 - 04/27/11 01:07 PM (12 years, 9 months ago) |
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Quote:
k00laid said: agar is for more than saving your cultures, OP..
I agree
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Omnicracker
Crusted Trolltivator


Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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To what degree?
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Omnicracker
Crusted Trolltivator


Registered: 09/18/05
Posts: 1,421
Loc: Wal-Merica
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only a degree
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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I see
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k00laid
NEMO


Registered: 05/03/10
Posts: 19,636
Last seen: 6 months, 5 days
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teehee
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Comprehension


Registered: 05/21/18
Posts: 87
Last seen: 4 years, 2 months
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Indeed
Tissue to Agar to LC
very high success rate
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 Legal Disclaimer! Everything I post is fictitious and mere role playing game as a scientist.
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