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OfflineJoelLoveland5
Giant Reptile
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Registered: 11/11/06
Posts: 222
Last seen: 16 years, 10 months
LC Media
    #6592523 - 02/21/07 01:57 AM (16 years, 11 months ago)

I recently bought a pre-sterilized LC media which I intended to clone with using a tek in which you stab the lower stem of a mushroom with a needle to get a core biopsy. You then inject the sample into the LC media.

Here is a link to that tek,
http://www.shroomery.org/forums/showflat.php/Number/5874305#5874305

I noticed in this tek creamcorn mentions that you should make the injection site hole in the lid slightly bigger than the needle itself in order to not create a vacuum.

This got me a little worried because I realized that the LC media I ordered had a self healing injection site and was pre vacuum sealed. I realized this because when I took the core biopsy and went to inject it, as soon as I entered the needle through to the inside of the jar it sucked the sample right out itself.

Was I correct to deviate from the tek or should I go and pop a hole in that thing and cover with some micropore tape as the tek suggests.

Later

-JL5000


--------------------
Newb Friendly Tek for The Lazy by "Hydro420"
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5278987/an/0/page/0

Great Newb Friendly Cloning Tek by "Creamcorn"
http://www.shroomery.org/forums/showflat.php/Number/5874305#5874305

One Goal, One Passion - Joel Loveland

"Dont Be Wise In Your Own Opinions"


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OfflineOatman2000
-=Outa Space=-
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Folding@home Statistics
Registered: 05/10/05
Posts: 2,877
Loc: Planetary Nebula
Last seen: 1 year, 4 months
Trusted Cultivator
Re: LC Media [Re: JoelLoveland5]
    #6592534 - 02/21/07 02:05 AM (16 years, 11 months ago)

vaccume happens... don't wory about it.

as long as it wasn't 12cc's of MS solution, your fine.

your clone is in there and will hopefully produce nice LC hyphae :thumbup:


--------------------
Spawning to COIR
:thumbup:  My Chocolate Recipe
WBS QUART SPAWN JAR PREPERATION
----------------------------

4-PO-DMT; 4-phosphoryloxy-N,N-dimethltryptamine


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OfflineJoelLoveland5
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Registered: 11/11/06
Posts: 222
Last seen: 16 years, 10 months
Re: LC Media [Re: Oatman2000]
    #6592558 - 02/21/07 02:20 AM (16 years, 11 months ago)

what is MS solution, I saw you refer to it in another post of mine too.

Tank you for the help

Later

-JL5000


--------------------
Newb Friendly Tek for The Lazy by "Hydro420"
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5278987/an/0/page/0

Great Newb Friendly Cloning Tek by "Creamcorn"
http://www.shroomery.org/forums/showflat.php/Number/5874305#5874305

One Goal, One Passion - Joel Loveland

"Dont Be Wise In Your Own Opinions"


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Invisibleagar
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Registered: 11/21/04
Posts: 9,056
Loc: Somewhere Else
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Re: LC Media [Re: JoelLoveland5]
    #6592624 - 02/21/07 03:10 AM (16 years, 11 months ago)

MS = MultiSpore


--------------------


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Invisiblecreamcorn
mad scientist
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Registered: 03/13/06
Posts: 2,962
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Re: LC Media [Re: agar]
    #6593343 - 02/21/07 11:10 AM (16 years, 11 months ago)

my concern with the vaccum is more or less on the opposite end when it comes time to use the colonized solution... the vaccum is going to fight you as you try to draw back on the syringe and suck up solution... it wants to pull your plunger down while you want to pull it the opposite way... well you get the picture. :wink:

a totally sealed jar is also not the greatest in a gas exchange scenario either, but the liquid itself should hopefully naturally contain enough dissolved gases to get you some growth.  (i.e. not optimal but not a deal-breaker for a successful LC either.)  i'd let it be though and cross those bridges when/if it comes.  might not be a problem.  wouldn't screw with it for now.


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OfflineJoelLoveland5
Giant Reptile
Male

Registered: 11/11/06
Posts: 222
Last seen: 16 years, 10 months
Re: LC Media [Re: creamcorn]
    #6593600 - 02/21/07 12:20 PM (16 years, 11 months ago)

Thanks creamcorn. As always extremely helpful

When it came time to do this I had a copy of your tek right in front of me. I was a little thrown off my game to find that the LC media I ordered had a self healing injection site.

In my defense I was also a little thrown off my game by the 6 pack of coronas (kinda slowed my thinking process and problem solving abilities).

Anyway, as I stated in another post I took about 4 samples from three seemingly closely related mushrooms and put them all in one LC media jar.

I am a little worried I might have contaminated it because of the large amount of samples taken, and I also forgot to wipe down the injection site with alcohol.

All my other utensils and environment were sterilized pretty meticulously, so I'm not that worried, guess I will just have to see how things go.

Long story short...
Dont drink and cultivate

By the way creamcorn, when I steam sterilized the syringe I was pressed for time. I did the whole 30 min of sterilization but then through the tinfoil cover syringe in the freezer. When I went to get it the syringe had a little ice in it. Could this hurt any part of the process?

-JL5000


--------------------
Newb Friendly Tek for The Lazy by "Hydro420"
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5278987/an/0/page/0

Great Newb Friendly Cloning Tek by "Creamcorn"
http://www.shroomery.org/forums/showflat.php/Number/5874305#5874305

One Goal, One Passion - Joel Loveland

"Dont Be Wise In Your Own Opinions"


Edited by JoelLoveland5 (02/21/07 12:46 PM)


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