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OfflineSeril
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Registered: 01/28/02
Posts: 246
Loc: PNW
Last seen: 11 years, 7 months
1st Liquid Culture, arrrgh!
    #616756 - 04/23/02 10:12 PM (15 years, 5 months ago)

4 tsp dextroe was added to a clean quart jar containing 400ml distilled water. (1-tsp:100-ml)
the jar was PC'd at 15psi for 15-20 minutes.
after cooling below 85F, 1cc of Equadorian spore solution was injected into the jar.
cap screwed on tight.
jar is then placed into incubator at 85F.

.....2 days later.... no sign of anything.. water still crystal clear.

1) do EQ's not germinate in dextrose?
2) can this guy use the SAME tek, but replace dextrose with honey to get germination from EQs?
3) can this guy now cut some pieces of mycellium from a cake he has fruiting, drop them into the jar, and expect a good colonization of the jar?

any info appriciated.
-seril


--------------------
Seril,
Political language -true of all political parties- is designed to make lies sound truthful and murder respectable, and to give solidity to pure wind..


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OfflineWorkmanV
Psilocybe Microscopist
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Re: 1st Liquid Culture, arrrgh! [Re: Seril]
    #616861 - 04/23/02 11:55 PM (15 years, 5 months ago)

Pure dextrose is missing nutrients required for growth. Try using recipes that work for agar media but just leave out the agar when making nutrient broths. PDYA (Potato Dextrose Yeast Agar) works good (note the addition of complex additives to the dextrose). You might also try MEA (Malt Extract Agar) using light tan malt from the brew store. Honey works sometimes but it isn't ideal. Just remember that mycelium needs a bit more than just sugar and water.


--------------------
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Offlinebadfish89
drunk by noonbut thats ok

Registered: 01/31/02
Posts: 45
Loc: phoenix
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Re: 1st Liquid Culture, arrrgh! [Re: Seril]
    #616961 - 04/24/02 02:40 AM (15 years, 5 months ago)

try using KARO or a like substance (light corn syrup) with the same ratio, 1 tsp per 100 ml. i recenlty did that using EQ's and they grew perfectly. so they can def. grow in Karo but I am not sure about dextrose, as I have not used it. you could also give it a couple more days......


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cant fight against the youth


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InvisibleSixTango
Mycota

Registered: 01/21/02
Posts: 1,996
Loc: A little North of Paradis...
Re: 1st Liquid Culture, arrrgh! [Re: badfish89]
    #617034 - 04/24/02 05:08 AM (15 years, 5 months ago)

My vote goes to 50/50 dry light malt & dextrose. They are available at any home brew supply store & cheap. At least it works for me. If it has gunk or carmalizes after PC'ing. Filter through a coffee filter & PC again. It should look the color of flat light beer. Mine do better with a filter to allow ample air exchange. Liquid culture is a contam magnet. It all has to be squeaky clean, or comtam city The upside is super fast colinization of whatever you put the myc juice to.
SixTango -- out


--------------------
~whiskey river rafting, hot tubbing, dirty dancing & spending money on - wild women - having fun & just gonna waste the rest~


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InvisibleSorted
Monkee
Registered: 12/27/98
Posts: 301
Loc: UK
Re: 1st Liquid Culture, arrrgh! [Re: SixTango]
    #617047 - 04/24/02 05:54 AM (15 years, 5 months ago)

Anyone know what the actual nutrient requirements for mushrooms are? I was wondering the other day if the yeast nutrient that homebrew shops sell could be added to a dextrose + water mix for liquid cultures..hmm


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Offlinesylo
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Registered: 10/30/00
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Re: 1st Liquid Culture, arrrgh! [Re: Seril]
    #617051 - 04/24/02 06:10 AM (15 years, 5 months ago)

I agree that dextrose alone has too few nutrients and contams are a big problem with liquid culture. Also, it takes three to five days for the spores to germinate. You can use a cake to spawn liquid but takes a sterile environment and good technique to make the transfer. A big problem is that spore solutions and cakes grown from spores are usually not Simon pure, absolutely one hundred percent contam free and this is what you need to start a liquid culture. The best way to go is to grow out the spores on agar and pick good mycelium colonies from the agar leaving any contams behind. I have found that the best way to grow in liquid culture is to keep the liquid shallow, no more than a half inch deep, and float a few tooth picks on top, AND NEVER SHAKE THE LIQUID. The mycelium grows as a tough, leathery layer across the surface and the tooth picks help keep it afloat. If the culture grows below the surface, you get a poor yield and a cottony mush that is hard to dry.


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Offlinecantankerous
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Registered: 04/05/02
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Re: 1st Liquid Culture, arrrgh! [Re: Seril]
    #617333 - 04/24/02 02:13 PM (15 years, 5 months ago)

Seril,

Pertaining to the last question you posted: I have taked anice piece of ryzimorphic growth from a cake and dropped it into a solution of water and about 1% h2o2 and shaken vigorously. This was then injected into a regular .5 pint jar with substrate. Within three days I started to see growth and after about ten days the jar was well past half done with VERY nice ryzimorphic growth all around. I would suggest a try at it if you have room for experimentation.

beatle


--------------------
peace,
beatle

"Friday is my high day"


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OfflineSeril
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Registered: 01/28/02
Posts: 246
Loc: PNW
Last seen: 11 years, 7 months
Re: 1st Liquid Culture, arrrgh! [Re: sylo]
    #617368 - 04/24/02 02:52 PM (15 years, 5 months ago)

thanks for all the helpful information folks. my friend went ahead and cut some pieces of mycelium from a cake in the fruiting chamber, and drop it in the jar as sterile and fast as possible. 12 hours since then, no sign of anything new, except a piece of floating mycelium (that my friend added added).
I'm willing to bet this is a dud, as predicted...
but where then, shall he use this McLarge 10lb bag of dextrose he has?

my friend is going to start agar work this weekend.
he'll be reading up. :tongue:
-seril
 


--------------------
Seril,
Political language -true of all political parties- is designed to make lies sound truthful and murder respectable, and to give solidity to pure wind..


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Offlinejuanell
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Male

Registered: 02/27/06
Posts: 7
Loc: Florida Flag
Last seen: 2 years, 4 months
Re: 1st Liquid Culture, arrrgh! [Re: Seril]
    #5350749 - 02/28/06 09:34 PM (11 years, 6 months ago)

ok i saw this wicked expencive stuff "Keepers Brew" what the deal i saw a few things on other sites but all have diffrent spore drop formatt, but "keeper" has this set up he stands by how can this liquid be substituteb but keeping his spore drop formatt at http://www.sporetradingpost.com/new_tech_brew.htm this is not an "add" honist question


--------------------
May i Mumble Dog Face in the Banana Patch?
Current Strains


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Invisibletiny_rabid_birds
Nocturnal
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Registered: 11/08/05
Posts: 15,653
Loc: estados unidos
Re: 1st Liquid Culture, arrrgh! [Re: juanell]
    #5350763 - 02/28/06 09:39 PM (11 years, 6 months ago)

What's up juanell, welcome to the shroomery!

Now, first thing's first... Keeper is a dick and you should avoid all his products at all cost.

Secondly, when you're using the search function or the related threads feature, make sure you check the dates before posting. This bad boy is nearly 4 years old.

Anyways, welcome to the shroomery, peace.


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OfflineLimelight_Liver
rushing on
Registered: 02/28/06
Posts: 26
Loc: outside the guilded cage
Last seen: 10 years, 9 months
Re: 1st Liquid Culture, arrrgh! [Re: badfish89]
    #5350980 - 02/28/06 10:35 PM (11 years, 6 months ago)

Karo works, it's not the absolute best, but it'll work.


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