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Offlinemycocozm


Registered: 05/13/10
Posts: 181
Last seen: 3 years, 25 days
Liquid culture trouble
    #14093699 - 03/09/11 05:14 PM (12 years, 10 months ago)

So I've been using the following recipe:

1 cup of water
2 tbsp light malt extract

90 min at 15 psi

Inoculate with H. ulmarius agar wedges, stir 15 minutes a day for 10 - 14 days.  It looks great by the end of this.  There are fragments floating everywhere, some floating on the surface and fuzzing out.  The agar wedge has become a blob but mixes thoroughly when stirred with the magnetic plate.  The solution is a nice amber color, like its supposed to be.  No visible contams. 

I stir this solution once more and backfill into a syringe, inject this into grain jars (same recipe that does fine when just receiving wedges) at about 15 mL per quart jar.  The bulk of the mycelium is still at the bottom of the jar with the agar wedge blob, it doesn't get sucked into the syringe, but the solution that does seems to be quite myceliated. 

But there is no growth after many days.  Nothing at all.  What am I missing?


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InvisibleDoc_T
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Registered: 03/06/09
Posts: 42,395
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Re: Liquid culture trouble [Re: mycocozm]
    #14093877 - 03/09/11 05:42 PM (12 years, 10 months ago)

15 mL per quart? Sheesh, try 1 or 2.

Also, stir it up. If the myc is heavy and settles, it can be tricky to shoot.

Why not do Grain LC?


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Invisibleslapphappypill
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Registered: 11/07/10
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Re: Liquid culture trouble [Re: Doc_T]
    #14093913 - 03/09/11 05:49 PM (12 years, 10 months ago)

I use about 4-5 CC's of LC in my qt jars, they colonize fast as well no prob. 4CC for a pint, and 2cc per 1/2 pint. currently my LC has colonized 90% of my wbs in 1.5 weeks without a shake yet....


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We think we have freedom, but we're all just mice in a maze....

                   

FYI: I stole all my pix off google! F+ PORN! Here is a shit-ton of porn by yours truly!

I have FINALLY written up a couple teks as to how SHP has done things in the past. DISCLAIMER: This is not for the newbie to mycology, and not going to work for everyone! This is simply what works for one person when other teks and methods have failed miserably!
~~~~~How SHP does their unconventional WBS Prep!! (NO DRY METHOD)~~~~
~~~~~SHP's highly disputed method of doing ALL their work outside of a flow hood or a Still air box!~~~~~
~~~~~Troubles harvesting the side and bottom pins in your mono? Learn how!! Dunking included ;-)~~~~~


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Offlinemycocozm


Registered: 05/13/10
Posts: 181
Last seen: 3 years, 25 days
Re: Liquid culture trouble [Re: Doc_T]
    #14093925 - 03/09/11 05:51 PM (12 years, 10 months ago)

Grain LC?  You mean starting the LC with grain?  Isn't that a little bass ackwards?


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Invisibleslapphappypill
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Registered: 11/07/10
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Re: Liquid culture trouble [Re: mycocozm]
    #14093942 - 03/09/11 05:54 PM (12 years, 10 months ago)

no, you make a small 1/2 pint with grain, inject spores, let it colonize, the inject sterile water and shake, suck it back up into the syringe and then noc up something with the mc water.

It takes the guess work out of whether or not you're lc is contamed as LC doesn't show visible contams


--------------------
We think we have freedom, but we're all just mice in a maze....

                   

FYI: I stole all my pix off google! F+ PORN! Here is a shit-ton of porn by yours truly!

I have FINALLY written up a couple teks as to how SHP has done things in the past. DISCLAIMER: This is not for the newbie to mycology, and not going to work for everyone! This is simply what works for one person when other teks and methods have failed miserably!
~~~~~How SHP does their unconventional WBS Prep!! (NO DRY METHOD)~~~~
~~~~~SHP's highly disputed method of doing ALL their work outside of a flow hood or a Still air box!~~~~~
~~~~~Troubles harvesting the side and bottom pins in your mono? Learn how!! Dunking included ;-)~~~~~


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InvisibleFungal growth
Lootinint
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Registered: 03/21/10
Posts: 3,641
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Re: Liquid culture trouble [Re: mycocozm]
    #14093956 - 03/09/11 05:58 PM (12 years, 10 months ago)

i may be wrong here, but i think your mix is too rich. if your lc is
too nutritious your myc wont want to jump to the less nutritious seed.
1 teaspoon of sugar is 4 grams. i only put about 2 grams lme in 1 qt jar.
Agar's recipe calls fir 1 tsp lme and 1 tsp dextrose in 1 qt water,
so theres room to play around with ratio's, but 2 tablespoons to 1 cup water is
WAY too much imo.


Edited by Fungal growth (03/09/11 05:59 PM)


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Offlinemycocozm


Registered: 05/13/10
Posts: 181
Last seen: 3 years, 25 days
Re: Liquid culture trouble [Re: Fungal growth]
    #14093983 - 03/09/11 06:02 PM (12 years, 10 months ago)

Thank you all, much help.


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