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OfflineYaMoonSun
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Kind of a dumb LC question
    #22167175 - 08/30/15 11:12 PM (8 years, 4 months ago)

So I took a tissue sample from a mushroom yesterday and made a liquid culture. I know it depends on the phenotype, but on average, how long does it generally take a culture to mature? :shrug:


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OfflinePussyFart
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22167206 - 08/30/15 11:23 PM (8 years, 4 months ago)

Quote:

YaMoonSun said:
So I took a tissue sample from a mushroom yesterday and made a liquid culture. I know it depends on the phenotype, but on average, how long does it generally take a culture to mature? :shrug:



What do u mean by "culture"? The LC? Maybe a week and a half to 2 weeks, then test jars/dishes should be knocked up and grown out to test viability and cleanliness.......


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Re: Kind of a dumb LC question [Re: PussyFart]
    #22167414 - 08/31/15 01:09 AM (8 years, 4 months ago)

You made a LC from raw tissue?? :shocked:


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Re: Kind of a dumb LC question [Re: PirateSwazey]
    #22168622 - 08/31/15 11:17 AM (8 years, 4 months ago)

Op if you make use of a stir plate with stir bar by weeks end you will have a nice culture going. After 2 weeks it will be dense. It is advised to stir using a stir plate a few times a week. Even a daily stir will benefit and expand growth. If your not using a stir plate it may take a little longer.


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OfflineYerow
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Re: Kind of a dumb LC question [Re: eatyualive]
    #22168669 - 08/31/15 11:25 AM (8 years, 4 months ago)

You cant take a pin/or tissue and drop into LC.. Ever tried dropping tissue, or a pin on agar? Tried watching what happens if you just let it grow and not do transfers? :shrug:


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OfflinePussyFart
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Re: Kind of a dumb LC question [Re: Yerow]
    #22168682 - 08/31/15 11:28 AM (8 years, 4 months ago)

Quote:

Yerow said:
You cant take a pin/or tissue and drop into LC..



Sure you can.....he just did....


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OfflineYerow
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Re: Kind of a dumb LC question [Re: PussyFart]
    #22168698 - 08/31/15 11:31 AM (8 years, 4 months ago)

But.. ffs lol


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Offlinetetherface
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Re: Kind of a dumb LC question [Re: Yerow]
    #22168728 - 08/31/15 11:38 AM (8 years, 4 months ago)

I use 9er tek macerated tissue clone juice to do most of my LC's with great success so as long as you did it in a SAB  with good sterile procedure you should be fine


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OfflineYerow
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Re: Kind of a dumb LC question [Re: tetherface]
    #22168746 - 08/31/15 11:42 AM (8 years, 4 months ago)

This makes no sense in my head, why on earth would anyone do this? You have no idea about genetics, traits or anything. No way you can take a pin or tissue and drop to LC even in front of a flowhood and not fuck it up. You gonna pick the pin or tissue from the substrate in 'sterile' condition? And then straight to LC? WTF :mindblown: :herpyderp: :justno: :kummeli: :feelsbadman:


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InvisibleMad Season
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Re: Kind of a dumb LC question [Re: Yerow]
    #22168764 - 08/31/15 11:48 AM (8 years, 4 months ago)

I agree it's bad techniques, and extremely risky for literally no reason. Agar is just as easy as LC. You just make any LC, and add 2g of agar per 100 ml.

That being said my first clone i did tissue to LC and got some amazing results. Beginners luck though because I tried again a few times after and failed miserably on almost all of them. Thus I do agar lol


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OfflineYerow
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Re: Kind of a dumb LC question [Re: Mad Season]
    #22168772 - 08/31/15 11:50 AM (8 years, 4 months ago)

Taking my hat off and shutting up


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InvisibleMikeBearPig
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Re: Kind of a dumb LC question [Re: eatyualive]
    #22168776 - 08/31/15 11:51 AM (8 years, 4 months ago)

Quote:

eatyualive said:
Op if you make use of a stir plate with stir bar by weeks end you will have a nice culture going. After 2 weeks it will be dense. It is advised to stir using a stir plate a few times a week. Even a daily stir will benefit and expand growth. If your not using a stir plate it may take a little longer.





I prefer not to stir mine at all.  It grow's just as fast with out.. with 1 major difference

When you stir, it can mask contaminations.  I have noticed if I don't stir and just let them do their thing, by 2 weeks all available nutrient's are gone and it is as big as its going to get.

However.. If you don't mix, it allows the contamination to keep feeding and grow fast.  Which is a good thing because it makes your contaminated cultures VERY easy to spot.

I only use MS honey liquid cultures, this method is the only thing that saves me from all the bullshit that comes with LC's.

Also, LC's with tissue samples are pretty much a 99% chance of failure, unless you have a flow hood.. Even then, it is a daunting task.

I've done well over 40+ attempts at cloning in all types of conditions in my SAB.  Only ever got 1 usable sample and it was shitty.  This is not to say you might have better luck, it's just something I would advice against.


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: PussyFart]
    #22168821 - 08/31/15 12:07 PM (8 years, 4 months ago)

Quote:

Mad Season said:
I agree it's bad techniques, and extremely risky for literally no reason. Agar is just as easy as LC. You just make any LC, and add 2g of agar per 100 ml.

That being said my first clone i did tissue to LC and got some amazing results. Beginners luck though because I tried again a few times after and failed miserably on almost all of them. Thus I do agar lol



If I had agar I would be using it, but for the time being, LC will have to work.

Quote:

eatyualive said:
Op if you make use of a stir plate with stir bar by weeks end you will have a nice culture going. After 2 weeks it will be dense. It is advised to stir using a stir plate a few times a week. Even a daily stir will benefit and expand growth. If your not using a stir plate it may take a little longer.



I've been giving it a little stir each day by rotating the jar in a circular motion. Looks good so far. No signs of bacteria that I can see.


Quote:

PussyFart said:
Quote:

Yerow said:
You cant take a pin/or tissue and drop into LC..



Sure you can.....he just did....



:lol: :cheers:


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22204121 - 09/07/15 07:54 PM (8 years, 4 months ago)

Look contaminated?
webm


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OfflineiSmkGrnBud
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22204253 - 09/07/15 08:19 PM (8 years, 4 months ago)

Hard to tell. It's LC. That's what sucks about LC. :shrug: All you can really do is use it and see what happens. Transfer some onto agar?


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: iSmkGrnBud]
    #22204274 - 09/07/15 08:22 PM (8 years, 4 months ago)

Did an inoculation today, going to wait a few days and see how it plays out.


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Re: Kind of a dumb LC question [Re: Yerow]
    #22204468 - 09/07/15 09:00 PM (8 years, 4 months ago)

I grew for over 10 years without agar.
I made clones in lc's. Take part of the mushroom and put in ziploc bag for a few days. When the myc starts getting fluffy, you can see if it has any obvious contams. Then drop it in an lc. Wait a week and bam.
How about this one?  Had a pin in a tub and dropped it in a jar of grain. Did some g2g and walla
http://www.shroomery.org/forums/showflat.php/Number/20964499


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: azur]
    #22206676 - 09/08/15 10:57 AM (8 years, 4 months ago)

Very much appreciating the tip :cheers:
Will give that a shot after there's some fruits on these new cakes. :super:


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OfflinePussyFart
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Re: Kind of a dumb LC question [Re: azur]
    #22206708 - 09/08/15 11:06 AM (8 years, 4 months ago)

Quote:

azur said:
How about this one?  Had a pin in a tub and dropped it in a jar of grain. Did some g2g and walla
http://www.shroomery.org/forums/showflat.php/Number/20964499



I have done this before.....10 pins, 10 jars, only one that didn't contam.....

This is when I learned that using something that has been sitting in open air is not good to just drop into a sterile grain jar.....


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Invisibleazur
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22206725 - 09/08/15 11:12 AM (8 years, 4 months ago)

Quote:

YaMoonSun said:
Very much appreciating the tip :cheers:
Will give that a shot after there's some fruits on these new cakes. :super:



I really recommend putting the pin in a ziploc bag for a few days first. Although agar is superior


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Re: Kind of a dumb LC question [Re: PussyFart]
    #22206729 - 09/08/15 11:13 AM (8 years, 4 months ago)

Quote:

PussyFart said:
Quote:

azur said:
How about this one?  Had a pin in a tub and dropped it in a jar of grain. Did some g2g and walla
http://www.shroomery.org/forums/showflat.php/Number/20964499



I have done this before.....10 pins, 10 jars, only one that didn't contam.....

This is when I learned that using something that has been sitting in open air is not good to just drop into a sterile grain jar.....



I was using a spare room strictly for tubs and was running a small hood in the room 24 Hours a day


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: azur]
    #22222374 - 09/11/15 02:45 PM (8 years, 4 months ago)

Inoculated like 8 times each, and not a single bit of growth anywhere. Perhaps I shouldn't try to incubate in 104 degree weather :facepalm3:


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OfflinetaGyo
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22222520 - 09/11/15 03:10 PM (8 years, 4 months ago)

:lol:, yeah that's pretty bad.


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: taGyo]
    #22224861 - 09/11/15 10:54 PM (8 years, 4 months ago)

So inoculate again, and leave them indoors.. :confused:


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OfflineiSmkGrnBud
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Re: Kind of a dumb LC question [Re: YaMoonSun]
    #22226099 - 09/12/15 08:17 AM (8 years, 4 months ago)

Quote:

YaMoonSun said:
So inoculate again, and leave them indoors.. :confused:




No I wouldn't trust that, not with them sitting in 104f. Best chance you have would be to re sterilize jars before you knock them up. I still wouldn't be too hopeful, you've probably lost a good bit of moisture content in those cakes. I wouldn't waste any spores on those jars. :shrug:


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OfflineYaMoonSun
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Re: Kind of a dumb LC question [Re: iSmkGrnBud]
    #22228974 - 09/12/15 08:54 PM (8 years, 4 months ago)

It's a 3cc liquid culture syringe


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