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Offlineecotopia
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Liquid culture question
    #19503940 - 01/31/14 11:07 PM (10 years, 11 months ago)

Trying to find an answer to this question and I'm struggling.

Is it possible to produce an isolate genotype LC?
If it can't be done directly then is it possible to isolate on agar and then use the mycelium from that to inoculate a LC?

I want to grow isolates for targeted genetics but I also want to use LC so I can add (hydrogen peroxide?) to the sub to fight off contams. I read that if you use LC then (hydrogen peroxide?) won't harm your mycelium.
Is this correct?

I wrote hydrogen peroxide as a question because I can't remember if this is in fact what I read


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OfflinePussyFart
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Re: Liquid culture question [Re: ecotopia]
    #19503954 - 01/31/14 11:10 PM (10 years, 11 months ago)

You cannot isolate in a LC.

Only on agar.

No peroxide, ever.

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Re: Liquid culture question [Re: ecotopia]
    #19503956 - 01/31/14 11:10 PM (10 years, 11 months ago)

skip the peroxide and yes do your isolating on agar first and knock your lc up with that or even just squirt some water on your dish (i use no pours for this) give it a shake and suck it back up, but keep in mind when you isolate they need to be tested before you go all out with them as do lc's in general so start with a couple cakes  or something


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Re: Liquid culture question [Re: ecotopia]
    #19503960 - 01/31/14 11:10 PM (10 years, 11 months ago)

you can't isolate in a LC, idk how you would go about doing that. You can get an isolate with agar and use it to inoculate a LC but if you have agar why bother with the LC in most cases.

Do yourself a favor throw the bottle of peroxide away, in the rare chance you get cobweb mold maybe try it. If you need to fight off bacteria you already did it wrong.

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Re: Liquid culture question [Re: ecotopia]
    #19503966 - 01/31/14 11:11 PM (10 years, 11 months ago)

Quote:

ecotopia said:


I wrote hydrogen peroxide as a question because I can't remember if this is in fact what I read



you probably did read it, so stop reading old shit lol, check tl's lc tek out


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Re: Liquid culture question [Re: cronicr]
    #19503986 - 01/31/14 11:15 PM (10 years, 11 months ago)

the thing with the shroomery is you can find anything you want to confirm your beliefs. There's 1000s of posts from all sorts of members a huge percentage of which know relatively little about mycology, even myself I consider to be amateur at best in mycology. You'll only see people post their success and rarely if ever their failures. So you're only seeing the people that got away, Hey I made this and this doing this and this and it for whatever reason worked the one time they did it and now they come here and say try this to a bunch of people. If you want to do yourself a favor look for TEKs that have a lot of views and replies less than two to three years old and see if people have success replicating them, find a grow along with a TEK and see how many people got results, not some kid saying h2o2 kills bacteria in a LC, I could go post that right now and half the people would believe it.

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Re: Liquid culture question [Re: bodhisatta]
    #19504044 - 01/31/14 11:26 PM (10 years, 11 months ago)

The only thing I would ever add to the LC is an antibiotic, such as ampicillin, kanamycin, chloramphenicol, etc. to reduce bacterial contaminants. Good sterile technique obviates the need for these though.


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Offlineecotopia
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Re: Liquid culture question [Re: cronicr]
    #19504087 - 01/31/14 11:35 PM (10 years, 11 months ago)

Ok so no peroxide. To be honest, using coir I didn't think that was a necessary step.
I just want to know I'm doing things right. Perfectionism is my affliction.

Right, so I can isolate on agar and then to LC but as bodhisatta said, why bother with LC if I have agar. Well without the need for peroxide (which is good news) that would be my first question.
Awesome, back to my original thought several weeks ago during the bulk of my research, isolate on agar, colonize isolate on rye grain, and spawn to coir substrate with gypsum, verm and maybe some chicken poo, but there appears to be mixed feelings on here about nitrogen and potency.
And I guess that's the final choice I need to make before I have my personal grow method

bodhisatta : Good and valid point you have there. I'm learning that as I go.
To be honest the method I'm working towards is coming together nicely and I have a lot of faith in my techniques once I've tried and tested and refined them. This is where perfectionism becomes a blessing, that mixed with tenacity and I get things right for me in the end. Much like my 6 month learning curve extracting a certain alkaloid out of a certain root bark. It has taken some refining but I'm getting great results now.

Like I said though, all I have to figure out is if I consider the nitrogen additive something worth believing in or not. I guess I could do myself a favour and do a side by side comparison, there would be no harm in trying as it's the only variable I have left. Everything else seems to fit snugly into place.

I look forward to the day I am the one giving out valuable advice on here and having some other valued member support the opposite opinion. :tongue:

P.s. I'm quite confident I have my sterile technique plans down. As I said I'm a tenacious perfectionist so that kinda obligates the good sterile technique.


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Re: Liquid culture question [Re: ecotopia]
    #19504099 - 01/31/14 11:37 PM (10 years, 11 months ago)

if your still gonna go the lc route pick up some whatman syringe filters


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Re: Liquid culture question [Re: cronicr]
    #19504113 - 01/31/14 11:42 PM (10 years, 11 months ago)

I wouldn't bother trying to isolate on agar yet unless you want to practice your sterile technique. I would do spores to agar then 1 to two transfers after that to get "limited" genetics the chances of it performing well is a bit better than a random isolate, if the isolate sucks then you waste your time but the limited ms grow is almost guaranteed to be at least average.

You can follow along in my grow along thread and read through it
and read through some of the pages to catch up, skimming or W/E

also skimming this thread

also feel free to send me a PM if you need agar help or some informative links

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Offlineecotopia
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Re: Liquid culture question [Re: bodhisatta]
    #19504141 - 01/31/14 11:48 PM (10 years, 11 months ago)

I understand that the agar culture will produce many different genotypes in the dish as it is MS and you can identify more valued genotypes by isolating a segment of mycelium which exhibits a more ropey and root-like texture rather than a fuzzy looking mycelium.
This way I can have a general idea about the characteristics of the fruit from the selected genotype to begin with, rather than waiting to see the fruit, isolating and failing.

Right?


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Re: Liquid culture question [Re: ecotopia]
    #19504151 - 01/31/14 11:51 PM (10 years, 11 months ago)

in theory, but not always. it's a good indicator but it's correctional not directly related. if you start one dish with spores it's going to look sort of gnarly with cottony growth take a good looking chunk or the furthest from the center chunk and put it to a new dish if it grows out and you see some ropy stuff take that and put it into your grain jar if it's clean or move it to a new dish and let that grow out and use that dish to inoculate a bunch of jars

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Re: Liquid culture question [Re: ecotopia]
    #19504157 - 01/31/14 11:52 PM (10 years, 11 months ago)

don't worry too much about cottony/rhizzo growth, that can be switched simply by adding/subtracting nutrition amounts in your agar but for the most part you won't even se any sectoring till arounr 4-5 transfers, so like bod said just shoot for clean growth first and clone when the time comes


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It doesn't matter what i think of you...all that matters is clean spawn
I'm tired do me a favor

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