Subtopic	Posts	Updated	Creator
	More Liquid Culture Discussion	85	12/19 11:33pm
	"Pure Culture"	-
	Honey water mycelium tek LIDS	19	12/14 02:21pm	I feel so weird
	Tell it to me straight	9	6/03	EvilShroomer
	Storing Liquid Culture	12	6/03	Peroxide
	Needle Gauge	10	11/28 11:40pm	plinkerdink420
	The Karo Tek Story	76	02/20 05:14am
	Liquid Inoculation, did I do it right?	7	12/18 03:24pm	jared
	Honey TEK Problem	4	12/25 06:54am	Sanchez

Ok, so I know this monkey who made successful honey water using Quote's Microwave Honey Water tek, with the PF strain. Now my question is, would this be considered superior breeding as opposed to straight spore injection? I realize colonization is faster, but is it comparable to the benifits of cloning a fruitbody?

By Nanook of the North (Nanook) on Wednesday, August 29, 2001 - 01:09 am:

You will not see as much genetic variation using liquid culture because you use far fewer Spores to start out with. Probably not a good idea if you are searching for something to clone... Still enough variation shows up that searching is generally... er.. fruitful.

Karo Tek or Dextrose excels if you intend to Clone. Inoculate the liquid media with sterile macerated mushroom tissue, it grows out in about 72 hours at 87*F. Then you have enough (storable) inoculum to make up syringes, Inoculate, and fruit a clone strain for quite some time.

By littlebro (Littlebro) on Monday, September 10, 2001 - 05:57 am:

has anyone had a problem using the honey tek, where the honey jar became contaminated and you didn't know it, then all the jars you innoculated went bad?

By Nanook of the North (Nanook) on Wednesday, October 10, 2001 - 09:34 pm:

Yeah, it has to be Sterile... You have to work in a Glovebox. Also, while many people do inoculate liquid cultures through a Polyfilled or taped hole in the lid, I have found the contam rates with these techniques are unacceptable. One contam here, and every jar you shoot is screwed.

By Liberty_Caps (Liberty_Caps) on Sunday, August 19, 2001 - 10:17 pm:

I was wondering if I do the honey tek syringes how long will these last? and will they colinize pf tek jars as fast, and as good as injecting actual spores? will they fruit as good or get the same amount of flushes?? smeems really easy but i dont grow large quanites so I dont want the ones I make go to wast I have lots of quistions on this subject so any response would be greatly appreacited ps. could i inject the honey water/spores into pf tek 1/2 pint jars that are colinizing really slow?

By Nanook of the North (Nanook) on Sunday, August 19, 2001 - 11:27 pm:

This looks like a fine place to post this.

Mycelial cultures in dextrose or honey last a good long time in the fridge. 3 months is not a problem on my stored cultures yet, I'd bet they last as long (refrigerated) as spore syringes. I'd say a year, not much (if any) more.

As far as the potency of mycelial solutions when used as inoculum for jars... You get a much faster start. A hot spore syringe takes three days to show growth for me, fresh mycelial solution shows visible growth in just 24 hours when jars are incubated at 87*F

You just can't lose with dextrose or honey teks to produce inoculum. This is the first tek a beginner would want to master in moving up to more advanced mycology: it's easy, it's cheap, it produces "hot" (fast and potent), storable inoculum. A single spore syringe can inoculate 750 jars in a week with a sugar (honey, dextrose, maltose, whatever) tek.

If your mycelial culture in sugar solution originated from spores, you will see genetic variation in your jars just as you would expect from a spore print or syringe, but you will likely see less genetic variation because you use fewer spores when you start out. The gene pool in your culture can be good, bad or indifferent... Depends on the dice.

Mycelial cultures in sugar solutions can also be started from sterile tissue samples taken from an outstanding fruit. In this way you can select from among the random genes displayed by spore germinations, and you can shoot jar after jar after jar of a proven clone.

You have to go back to spores at some point because there is a "biological clock" on the number of generations cloned mycelial will remain aggressive and fertile, but there is no reason you could not start a large number of dextrose jars from the original tissue sample and store these "master" cultures in the fridge. Regardless of scale, it's a great technique to improve productivity. Once you learn dextrose (glove box, sterile technique and the rest)... Your next step is learning to clone.

Nan

By Liberty_Caps (Liberty_Caps) on Monday, August 20, 2001 - 12:18 am:

alright thanx for the reply nanook,,, I was also thinking of making the honey tek but let the mycelium grow as much as it would then drain the water and eat the mycelium just a thought don't know if I could get enought to do the job tho?

By Nanook of the North (Nanook) on Sunday, September 30, 2001 - 03:38 am:

I read about a commercial operation once using liquid substrate (potato water) in five gallon carboys. Micron filters on aquarium pumps were used to bubble air through the bottles and mycellia was grown out. Once the bottles were fully colonized they were dumped out into a filter, the mycellium was pressed into cakes and dried.

This operation bypassed the fruiting stage and produced large yeilds according to the author, but the equipment requirements were a little extreme (and required a commercial sized pressure cooker to hold a 5 gallon carboy)... So, having seen the photos I know it is possible, even on a smaller scale, but I would miss seeing the fruits.

Nan

By Glassman (Glassman) on Sunday, September 16, 2001 - 01:21 pm:

I'm trying the honey tek from this board's teks for growing out mycellium for innoculation. I've got a nice thick pool of mucus looking mycellium, but I can't draw it up throught the syringe. It's too thick. Anyone have experience with this method. what is my next alternative for innocculating pf jars with this mycelium.

By Fishy1 (Fishy1) on Sunday, September 16, 2001 - 04:11 pm:

I make my own needles/ adapters out of aluminum on my lathe. You could drill out the plastic needle adapter and glue on a length of alum. tube as a big bore needle.
If I have time in the next week, I will make up a couple of the needles for a giveaway/contest or sumthin'. They work great! fishy1

By Glassman (Glassman) on Monday, September 17, 2001 - 12:51 am:

Wow, that sure would be awesome. in the meantime, does anyone have any suggestions of how to use the current mycelium I have?

By Nan (Nanook) on Saturday, October 13, 2001 - 01:07 am:

Purge: I use broken jar glass in the bottom of my dextrose jars before sterilizing. If you shake them occasionaly it breaks up the clumps and gives a nice suspension for shooting.
If it is already clumped you can whip it up with the tip of the syringe needle for a few seconds before drawing it up.

By brandeen on Friday, August 31, 2001 - 01:15 am:

First off, I love this place. Makes you feel warm inside. Couple q's regarding honey / dextrose tek.

If you were to grow out the tek as much as possible, how long until the sugar is used up (at 80 degree incubation), how much dried mycelium would we be talking about (after straining and drying), and is the mycelium potency relatively close to the fruitbody potency?

By the way, we don't have to expose the incubating jar to light do we? Also, does the mycelium grow on the bottom or top of the broth?

Woah! Sorry about all the q's, I suppose I'm a curious cat. Peace, bros.

By Karna (Karna) on Friday, August 31, 2001 - 11:20 pm:

How long it takes for the sugar to be used up is a difficult question. Colonization is faster the more spores/mycelium you inject. If I remember correctly, the maximum growth takes place a few days after the last of the sugars is used up. A good general rule is to check every day and stop incubation the day you see the most difference from the previous one. I would say with spores about 8-12 days.
No idea how much the strained dried mycelium would weigh. Not very much. It's not worth the effort from a potency point of view. It would take a lot of jars to get close to the effects off of say 2gms dried fruit.
Also, no you don't have to expose the incubating jar to light and the mycelium grows on the bottom of the jar with the occassionaly floater appearing.

By wang on Sunday, August 26, 2001 - 08:58 pm:

hi guys I still suck and havnt quite figured out the liquid culture...soooo can anyone tell whats up with my water? I used 1tsp to 75cc water,pc for 30 at 15 psi and they all have a yellow tint. is this normal? thanks
-wang

By Nanook of the North (Nanook) on Sunday, August 26, 2001 - 11:53 pm:

It is possible you had some Carmelization if the jars were sitting directly on the bottom of the pc when you sterilized or you heated at too high a setting. Rusty Lids will drip staining water inside dextrose jars during sterilization sometimes.

Jars are reusable, Karo is cheap. I would go back and check on why the solution is tinted, it should be crystal clear colorless. Reduce your heat, make sure the jars are not sitting on the bottom of the sterilizer.

Nan

By Nuecrew (Nuecrew) on Wednesday, October 03, 2001 - 01:12 pm:

Has anyone used mycelia water to innoculate 1/2pint jars like we use with spores.(PF Tek) Also two times now I've had very little mycelia growth after using mycelia water. I tried to be careful not to get lysol anywhere near it the last time. (9er Tek)Just need some feedback if anyone has gone down this road.

By Karna (Karna) on Friday, October 05, 2001 - 09:19 pm:

i use only liquid mycelium. Great results every time. Faster than spores. Make sure you grow it out till the mycelium reaches at halfway up the jar from the bottom. You'll know what I mean as it grows. That way there will be plenty of it which means more plentiful, faster growth.

By monkeyod (Monkeyod) on Friday, October 05, 2001 - 09:53 pm:

I'm running a few jars (1/2 pint) from a syringe and somemore started from mycelium. The myc. were started 1 week behind the spore jars and have almost caught up to them. Plus I got 4 syringes from about 1cc from a spore syringe...not to bad. I never have used Lysol, just clean the area you're going to be working in with Bleach/Water and go for it.

By Karna (Karna) on Friday, October 05, 2001 - 10:11 pm:

just as a reference following monkeyod's post here's a method known to do good just to illustrate that the potential (informational purposes only) for exponential expansion.

1cc of spores into each of four pint jars filled halway (200-250cc)with dextrose water or other culture medium
cultured till mycelium has reached almost till the surface (about 2 weeks or just under)
= 20 to 25 10cc syringes per jar
= at 1cc per jar that's 10 jars per syringe
= 200 to 250 colonized jars per jar of mycelium water.
This quantity leaves you free to change both sides of the equation according to your needs or patience.
For eg., at 2.5 cc per jar, we get 4 jars per syringe = 80 to 100 jars per myc jar but faster colonization.
Now, if you employ any one of a number of expansion, or "bulk" teks at, say 1:5 ratio (conservative), each colonized jar of substrate gives rise to 5 colonized jars worth = 1000 to 1125 colonized jars worth (1cc)
or 400 to 500 colonized jars worth (2.5cc) - from one jar of liquid culture.
Mind you that is just expansion at level one (one "bulk" spawn run)
You get the idea.

By Nanook of the North (Nanook) on Thursday, October 11, 2001 - 04:00 pm:

Wow...

>Now, can the mycelial water in your spore syringe be used to inoculate cakes?

Yes, that is it's intended purpose.

>Is this better than cloning?

Mycelial solution can be made from spores or cloned tissue. If you are cloning it beats the hell out of agar work hands down. If you are working with spores it will stretch out your supply a long, long way.

> Better than collecting spores from a mature shroom?

As long as your spores are clean (sterile) it makes no difference where the spores come from.

> Better than ordering a syringe from PF?

Go ahead and buy a syringe from PF, it's a good source of sterile spores. But using that syringe to inoculate a jar of honey or dextrose solution can produce dozens of syringes of mycelial solution capable of inoculating hundreds of jars.

> Would this mycelial water be better to use in the BRF/Verm mixture when making the cakes?

It is inoculum, fill sterile syringes with the solution and use it just like a spore syringe.

> Would it be better to dunk cakes in mycelial water?

No, and you would not dunk cakes in spore water either.

By Joe Mamma (Madscientist) on Friday, September 28, 2001 - 10:06 pm:

Can anyone tell me what to look for in contaminants in liquid mycillium? I have two jars. They both are mainly long strands of fuzzy white fungi. The longer ones tends to have a little blackness to them. Is this bad fungi or is this the good stuff? Any expert help would be appreciated.

By Lichen (Lichen) on Saturday, September 29, 2001 - 02:31 pm:

the rule of thumb is ANY color besides white indicates bacterial contamination. Mycelium is white--no exceptions. If you have black or pastel colors, throw it away without exposing yourself to the stuff. It can be dangerous.

By Brad (Raze) on Monday, November 19, 2001 - 01:46 pm:

Um, my dextrose jar germinated wednesday, as of now it has three fluffballs of spores that germinated, plus a bunch of other white fluffs floating around.
The honey, which I innoculated way b4 the dextrose germinated yesterday, it already has a 1cm layer off fluff on the bottom, I'd say its 3x as far ahead as the dextrose?

By quote: (Quote) on Monday, November 19, 2001 - 01:54 pm:

what's the question ?

By Brad (Raze) on Monday, November 19, 2001 - 07:17 pm:

I kind of messed up the second paragraph there... Its supposed to say that I started this dextrose jar ahead of the honey, by about a week. But yet the honey is just zooming along, while the dextrose jar only has three small balls of mycillium.... Why is this?

By Lichen (Lichen) on Monday, November 19, 2001 - 08:26 pm:

good question...my Karo jar attempts are struggling along...slow. My first contammed, so I'm trying another; my honey failed (I think) and I threw it out. My second Karo is incubating now. I don't have very many spores, let myself get behind, and my shrooms are down to TWO cakes in the terrerium, so I'm limited in my experiments right now. That's what I get for slacking off...But! I have a whole bunch of jars growing, some near birth, and so in a couple weeks I'll be a bit more up to speed on liquid culture and spore printing.

By Brad (Raze) on Monday, November 19, 2001 - 09:30 pm:

Is it ok to use liquid culture to start another liquid culture? Or woudl that have the same effect as cloning (degeneration of the DNA over time).
I have 2 syringes left, one more with 1CC in it. My friend just ordered 3 B+ syringes, we were gonna swap 1 for 1. So I only really have one Matias Romero to spare after this 1cc

By Karna (Karna) on Monday, November 19, 2001 - 10:35 pm:

Yes it is ok to use liquid culture to start another one.
Your dextrose jar could be behind for many reasons
a. the sugar content is not optimal or some sugars caramelized.
b. your dextrose source was impure.
c. The mycelium is fighting some intruders
d. you injected a lot more spores in the honey jar
whatever it is, I'd use the fastest grower to inoculate.

By Nan (Nanook) on Monday, November 19, 2001 - 10:49 pm:

Most definately. If you have a choice use the fastest, healthiest culture source possible to inoculate.

By Brad (Raze) on Tuesday, November 20, 2001 - 01:29 am:

I used 1cc for both...
I followed Nan's formula, 1tsp for 75CC water. Dextrose for wine & beer making.

By An guy (Boomer) on Tuesday, November 20, 2001 - 01:32 am:

As a nutrition source, honey is far more complete than dextrose. Shrooms and folks is different animals, yes, but think about it-

How long would you last on honey, as opposed to refined dextrose without becoming diseased?

I know Karo works, I am using it myself, but I am going to go to honey- Nan could tell you there's a lot more good stuff in honey than in anything refined, and shrooms are a life form.

I could be wrong here, and Karna's reasons or someone elses might be what's going on with your jars, I dont' know. This is just me thinking, along with this thought

Dextrose is a refined product, as is white rice. If that's all one eats, one starts getting all kinds of illnesses, as has been scientifically shown time and again.

Brown rice is far more complete as a food, as is honey.

If shrooms grew as well on white rice as they do on brown, reckon we'd some of us be growing on white rice, given that we try to use and recommend cheap, easy to get stuff and white rice is cheaper and more readily available than brown, generally speaking.

Dextrose, Karo, they work. And for some folk, who can't get good honey, ie, can only get the blended, cheap-ass, cooked, highly processed honeys, well, other sugars are as good as that.

But if you can get prime honey, well, I can, and that's what I'm gonna be using in the future.

So even if I'm wrong, I'm betting it was the honey.

By Lichen (Lichen) on Tuesday, November 20, 2001 - 01:43 am:

sound, in my opinion. Honey is good stuff.

By Nan (Nanook) on Tuesday, November 20, 2001 - 03:05 am:

No arguement here. The longer you grow in sterile culture, the more complete the nutrition must be.

3-10 days (Incubated) in 4% Dextrose is an efficiency practice.

Flash germination of spores or culturing of cloned mycelia is what it is about. Spores start germinating in within 24 hours when incubated (you can see them), you can Shoot thrashed dextrose in days when fired up with a PF syringe in a glovebox.

Honey is certainly more complex. If you intend to grow in liquid culture without shooting quickly... I would recommend Karna's Liquid Culture Media. It only makes sense.

But if you want something crystal clear, a simple sugar is all you need for a few days. Don't expect masses of mycelium, all you need is a "puff", thrash it and shoot it. Toss the remainder in the fridge. You don't want to grow mycelia out with Liquid Teks... You just want to get a quick germination or recovery from maceration, thrash it, and shoot it.

You can always shoot a few cc's into a richer media... Agar, PF Jars, whatever.

By Nan (Nanook) on Tuesday, November 20, 2001 - 03:13 am:

Ooops... It just occured to me that the Peroxidated Agar so loved by the solid inoculation tek people... Is pure Maltose solid culture.

By Brad (Raze) on Tuesday, November 20, 2001 - 03:59 am:

So you're saying the two puffs of mycillium in my dextrsose and 6 puffs in my honey are what I'm looking for? I can go now?

By Nan (Nanook) on Tuesday, November 20, 2001 - 04:15 am:

Yeah. Move the jars into the glovebox, open them, get a sterile needle and thrash and whip the puffs of mycelia. It does not hurt to suck them in and out of the syringe to fragment the puffs completely. Suck up a syringe of the fragmented mycelial culture and hold it to the light. You will see white tissue "floaties" suspended in the syringe. Make sure there is an air bubble in the syringe and keep the solution agitated.

Shoot what you need, seal the jar, pop it in the fridge, and you can shoot the rest at your leasure.

By Brad (Raze) on Tuesday, November 20, 2001 - 04:32 am:

I dont have a glovebox... and I have no where to store one. An incubator and fruiting chamber is already 2 rubbermaids under my desk that didnt used to be there. If I have one with two holes and gloves in it beside it, some suspicion may arise

By Nan (Nanook) on Tuesday, November 20, 2001 - 05:33 am:

So you are at least using Oven Tek?

By Brad (Raze) on Tuesday, November 20, 2001 - 05:35 am:

No... I use a room nuked with lysol

By Nan (Nanook) on Tuesday, November 20, 2001 - 05:58 am:

Hummm we need a stealth Glovebox design. A folding cardboard box set up 3-4 feet above the ground would be a great starter.

A Cardboard Glovebox and playtex gloves is better than Oven Tek imo.

Sheath plastic construction would be ideal with smooth Sanitizable surfaces. But make it so you can fold it up and tuck it away somewhere. It saves a lot of lysol and smell, and you get better sterile conditions (and focus) inside a box.

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