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Offlineshane4050
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Registered: 09/25/08
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Karo LC *pic*
    #21963910 - 07/19/15 09:10 AM (8 years, 6 months ago)

Made this LC up about 6 days ago, didn't put any glass or marbles in it and just used my microwave to prep/sterilize instead of messing with my pressure cooker. Used about .5cc spore solution Burma.



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Offlinenooberst
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Re: Karo LC *pic* [Re: shane4050]
    #21964157 - 07/19/15 10:14 AM (8 years, 6 months ago)

That's pretty risky using spores to lc


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OfflineMi-Go
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Re: Karo LC *pic* [Re: nooberst]
    #21964183 - 07/19/15 10:26 AM (8 years, 6 months ago)

How come? I plan on doing this with penis envy, all I ever get are ms spores I could never find a way to get lcs, I get from a sponsor too, they don't carry lcs for the desired strain.


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Invisiblemaddchef
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Re: Karo LC *pic* [Re: Mi-Go]
    #21964204 - 07/19/15 10:30 AM (8 years, 6 months ago)

Especially don't do this with PE from spores.


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In the land of the blind, the one eyed man is king.

All mushrooms are edible, but some only once.....                     

                                        Easier than cakes

I do science and shit.

:havesomescience:


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InvisiblePastywhyteMDiscord
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Re: Karo LC *pic* [Re: maddchef]
    #21964252 - 07/19/15 10:40 AM (8 years, 6 months ago)

Spore syringes are made from prints. Prints are made from fruits fruited in open air. Prints are pretty much impossible to get 100% aseptic.  Which means syringes are almost never 100% aseptic.

When a person uses a spore syringe to inoculate grains or cakes they are playing the percentages. You use a minimal amount of solution in the hopes that if there is a random mold spore you can spot the jar it ended up in and only lose that one. Bacteria does not usually do well on brf so many times people fruit through it and don't even know its there. But in LC that one mold spore will become a huge colony. That tiny amount of bacteria becomes a bloom. Because you're expanding the inoculate you are now contaming far more grains than you would with just the spores.

Spores to LC can work. But if they don't the fail is epic.

PE is likely to have bacteria mixed in with the spores because of the way the syringes have to be made. I would never attempt it with an LC ever.


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InvisiblebodhisattaMDiscordReddit
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Re: Karo LC *pic* [Re: Pastywhyte]
    #21964310 - 07/19/15 10:49 AM (8 years, 6 months ago)

Indeed one drop of spore solution may have 100s to 1000s of spores, a single small drop, but it may have 1 or 2 bacteria. The more drops of spore solution you use the much higher the chance of one of the few inevitable bacterium get in your LC. It only takes one bacterium to spoil the whole LC. Since a LC is liquid media unlike the 2D media agar the bacteria even if non motile will fuck the whole LC no problem. Most bacteria  won't make any smell or turbidity or anything easy to notice either. So you'll want to test it before using it on something important.


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Offlineshane4050
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Re: Karo LC *pic* [Re: bodhisatta]
    #21964611 - 07/19/15 11:50 AM (8 years, 6 months ago)

I figured it was risky but I had a tiny bit left in the syringe and figured mine as well, though when I made the jar some black specs of print entered from the syringe and two pools of myc have now grown around each one. Pretty rapidly I may say too. Hopefully I will get a 8/10 success ratio on my grain jars next week. Either way no loss to me as I have a popcorn jar of Burma I plan to g2g as well.


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Offlinekushroom
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Re: Karo LC *pic* [Re: bodhisatta]
    #21964677 - 07/19/15 12:12 PM (8 years, 6 months ago)

Away around the contam tho is easy, shoot spores onto agar, agar wedge to lc, wala should be clean as long as your sterile tek is good.


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:/



All submitted posts are by Someone Who Isn't Me (SWIM) -  and in any event are works of pure fiction or outright lies.  Any information, statement, or assertion contained therein should be considered pure unadulterated fictitious lies.


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OfflineMi-Go
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Re: Karo LC *pic* [Re: kushroom]
    #21965055 - 07/19/15 01:38 PM (8 years, 6 months ago)

can i do that with pes? Im trying to make the best out of that syringe man i want a big fuckin flush. I have some hawaiins PES too


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OfflineMi-Go
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Re: Karo LC *pic* [Re: Mi-Go]
    #21965064 - 07/19/15 01:40 PM (8 years, 6 months ago)

I want to work with liquid cultures, seems like the way to go, just dont have a mag stirrer.


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InvisiblePastywhyteMDiscord
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Re: Karo LC *pic* [Re: Mi-Go]
    #21965109 - 07/19/15 01:50 PM (8 years, 6 months ago)

You need to learn proper sterile tek before you can do LC. Otherwise expect a lot of fail.


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OfflineMi-Go
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Re: Karo LC *pic* [Re: Pastywhyte]
    #21965215 - 07/19/15 02:08 PM (8 years, 6 months ago)

Im guessing this is a good start?http://www.shroomery.org/forums/showflat.php/Number/21032261/fpart/1/vc/1
I live in an apt. Dont have a lab or flowhood at my disposal.


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InvisibleToadstool5
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Re: Karo LC *pic* [Re: Pastywhyte]
    #21965272 - 07/19/15 02:15 PM (8 years, 6 months ago)

How weird! I just started some honey water LC jars from some 3ml MS syringes (boredom). So far no sign of visible contams but i used slightly more honey ~8% and only one jar has started growth. If they continue to stall for two more weeks i will assume bacteria got them or the sugar was too much. If they start up soon and thrive perhaps the sugar content is what enabled me to stall the bacteria long enough for the mycelium to take over.

I do stupid shit sometimes :rofl:

Quote:

live in an apt. Dont have a lab or flowhood at my disposal.




I use a SAB in a bathroom with no issues. You have to read a lot, learn proper use of disinfectants, how to control air currents, and proper teks. Once you have more experience and learn more it will become easy to work cleanly.


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If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

AMU Teks :mushroom2: Stro's Write Ups


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Offlineshane4050
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Re: Karo LC *pic* [Re: Toadstool5]
    #21965889 - 07/19/15 03:44 PM (8 years, 6 months ago)

Yup a still air box can be built cheap, 5-10 bucks. Most people will have the materials around the house. And a bottle of Lysol spray in the smallest room of your house and work in there. Either way trial and error.


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Offlineblackout
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Re: Karo LC *pic* [Re: Toadstool5]
    #21966017 - 07/19/15 04:06 PM (8 years, 6 months ago)

Quote:

Toadstool5 said:
How weird! I just started some honey water LC jars from some 3ml MS syringes (boredom). So far no sign of visible contams but i used slightly more honey ~8%



next time try a lot less like 0.1-0.5% sugars.


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InvisibleToadstool5
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Re: Karo LC *pic* [Re: blackout]
    #21966107 - 07/19/15 04:28 PM (8 years, 6 months ago)

I would expect bacteria to take it over if i did them at such a low concentration. Im trying to see if i can get growth without bacteria by intentionally running them at 8%. Honey is possibly a mild biocide so im adding enough to eliminate all the bacteria and most but not all of the spores (i expect something will arise from using MS). It will probably fail but i like to try new things.

If i wanted clean LC i would simply isolate/clean on agar and then make a slurry of the agar/mycelium for inoculation.

On a side note, im mostly worried about zygosaccharomyces but i think they would bubble if present from the fermentation.


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If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

AMU Teks :mushroom2: Stro's Write Ups


Edited by Toadstool5 (07/19/15 05:01 PM)


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Offlineinvitro

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Re: Karo LC *pic* [Re: Toadstool5]
    #21966805 - 07/19/15 07:30 PM (8 years, 6 months ago)

Interesting concept,


Here are two charts showing MIC (minimum inhibitory concentration) of different types of honey.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3609166/figure/apjtb-01-02-154-g003/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3609166/figure/apjtb-01-02-154-g002/

Main article: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3609166/

It appears at first glance that the anti-microbial properties of most types of honey at low concentrations are due to h202, and pcing the LC has probably killed that aspect of your honey. 

The odds look stacked against this experiment, but good luck!


Edited by invitro (07/19/15 07:32 PM)


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InvisibleToadstool5
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Re: Karo LC *pic* [Re: invitro]
    #21967907 - 07/19/15 10:51 PM (8 years, 6 months ago)

Yeah the peroxidases are essentially gone but even non-peroxidase honey has also shown antimicrobial effects so god only knows what will happen, its interesting because i used a large amount of  inoculate.

Quote:

. However, another kind of honey, called non-peroxide honey (viz., manuka honey), displays significant antibacterial effects even when the hydrogen peroxide activity is blocked. Its mechanism may be related to the low pH level of honey and its high sugar content (high osmolarity) that is enough to hinder the growth of microbes. The medical grade honeys have potent in vitro bactericidal activity against antibiotic-resistant bacteria causing several life-threatening infections to humans. But, there is a large variation in the antimicrobial activity of some natural honeys, which is due to spatial and temporal variation in sources of nectar.




Im using my friends home grown honey, made with any nectar they can find within a few miles of his house so i really have no clue what will happen. :lol:

So far there is zero growth out of 7, 100ml jars except one with radial mycelium about 1cm in radius. Its been  about 5 days now :shrug:

It will be awesome if i can get a clean LC made from a MS syringe but i doubt i will. Im going inoc a dyed agar plate with it when its done to see if it is actually clean.


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If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

AMU Teks :mushroom2: Stro's Write Ups


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