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HypnotoadCroaked
Retired, but will check MSGs

Registered: 01/05/13
Posts: 1,168
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Clone from tissue culture. New to study.
#19175511 - 11/22/13 04:25 PM (10 years, 2 months ago) |
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I cloned a monstrous fruit and managed to make two out of three dishes without contamination. These two dishes are extremely rizo on the media I am using. I have made quite a few "MS" agar transfers, in order to create the fruit that I am cloning.
In the last 8 days it has blossomed to cover the entire bottom of a two cup PP5.
It is not an isolation, but I would like to preserve the clone in its entirety. Is this a practice? If the culture is deemed "clean" would a cut from the original fruit be a good option?
What is the best way to move clone to say 5+ different jars and still preserve the unique aspect of the fruit?
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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its a crap shoot as far as my understanding goes. that clone could have many many dikaryotic mycelium making it up, and when u clone, you get all of them. if u take that grown out plate from the clone and use it to inoculate some grain jars, without isolating, you will indeed have jars that represent the clones mycelial makeup, but the fruits that come will not necessarily be like the clone. I dunno if this makes sense, but its how I think of it... you clone a fruit on petri. it has myc A, myc B, myc C, and myc D (for simplicity sake) you use that to noc up grain, it grows out and u spawn it. fruits grow, those fruits don't necessarily have all A,B, C, and D myc, one fruit might have A and C and D, and another fruit might have B and D and another fruit might have B C and D. It is better than doing an MS grow because you have limited the number of genetics going into fruit body formation, but there will still be some diversity, which may not give you the qualities you are seeking in each of ur fruit bodies, but it will get you one step closer to where u wanna be. I take spores, noc up agar and do some transfers. When a plate looks healthy, but not necessarily isolated, I noc up grain and grow it out, fruit it, then take clone samples and isolate those further, because I know at least they are potential fruit forming specimens.
I don't know if preserving a clone is common practice, other than an isolated clone. Try it and see! Hope that helped.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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karp


Registered: 10/21/13
Posts: 310
Last seen: 8 years, 7 months
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if you want to colonize a bunch of jars from a clone add it to one jar, then let it fully colonize. next add some of the grains from the colonized jar to a bunch of other jars.
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Clone from tissue culture. New to study. [Re: karp]
#19175635 - 11/22/13 05:08 PM (10 years, 2 months ago) |
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be careful just dropping a clone in a sterile jar. its risky, but do it if u wanna take that risk. the avenue you have chosen is a safer bet, u can spot any contam that may have been part of the clone much better on a 2-d petri than a 3-d jar. u dunno what may have landed on a pin/fruit during its growth cycle. u can noc up a jar with growth from a clone if its on a clean plate.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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HypnotoadCroaked
Retired, but will check MSGs

Registered: 01/05/13
Posts: 1,168
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Re: Clone from tissue culture. New to study. [Re: karp]
#19179555 - 11/23/13 04:24 PM (10 years, 2 months ago) |
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Quote:
blindingleaf said:Hope that helped.
Yepper. That has cleared my questions up 100%. So using the entire genetic spread, I will simply be spawning a mixed bag of genetics still, but greatly limited versus spore inoculations. Thanks for your help!
Quote:
karp said: if you want to colonize a bunch of jars from a clone add it to one jar, then let it fully colonize. next add some of the grains from the colonized jar to a bunch of other jars.
I have been using agar to isolate clean growth away from possible contamination, AND then spawning clean cultures to grain. IMHO the best decision I made in this hobby is to start using agar. Though I am quite limited in my knowledge, agar is vastly superior to syringes full of mystery.
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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yea dude agar is the shit! only problem I have with it is when I have 4 different strains and forget to label them correctly.... so now I have 4 grows and I only know what 1 of them is haha
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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HypnotoadCroaked
Retired, but will check MSGs

Registered: 01/05/13
Posts: 1,168
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Re: Clone from tissue culture. New to study. [Re: blindingleaf]
#19179637 - 11/23/13 04:48 PM (10 years, 2 months ago) |
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Quote:
blindingleaf said: yea dude agar is the shit! only problem I have with it is when I have 4 different strains and forget to label them correctly.... so now I have 4 grows and I only know what 1 of them is haha
I am a bit fussy about making 100% sure of labels. With the amount of jars/tubs/bottles that can float from time to time it is imperative for me. I forget what I did yesterday let alone what is in the ball jar with the white plastic lid lol. I don't want to confuse oysters with something more sinister
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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yea I label the petris now. when I first started, I labeled the tupperware they were in, 2 stack of strains in each tupperware, and the lid of tupperware was labeled. one time, I took the lid off, and since the lid can go back on 2 different ways, I wasn't sure which way was correct, so I ended up with mystery petris.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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