Mr. B (Argonaut)
|Posted on Tuesday, December 11, 2001 - 03:42 pm:||
This is a Honey jar. Knocked up on 12-08
Mr. B (Argonaut)
|Posted on Tuesday, December 11, 2001 - 03:46 pm:||
This is Karo Tek knocked up on 12-8 also.
|Posted on Friday, November 02, 2001 - 04:09 am:||
Quoting Lichen on his three day old Honey Tek jar: "as of what--three days, now? it is developing a yellowish sort of stuff in the bottom....looks like sediment. I shake it up and it swirls around like a snowstorm paperweight, then settles again."
My reply is:
"See this is the problem with honey. Honey contains "hidden" impurities: wax and protein. When you cook honey the protein denatures, just like cooking the whites of an egg. The protein is not visible in fresh honey, cook it and the protein denatures and coagulates... Hence sediment. The wax, which is emulsified in fresh honey, separates out upon heating and forms scum... Some of it binds to the coagulated protein, some of it floats to the top. Then there is the color, which varies from honey to honey. It all makes it difficult for people new to liquid culture to determine off colors and contams. I am rapidly becoming motivated to do some experimental research with clear Karo syrup, which is available right on the shelf of any decent grocer... "
[ed. note: turns out a mycotopiate name "Purge" was the actual fellow who gave nan this idea, but as usual, nan neglects to credit his source, so i'll do it for him.]
And so I did. Here is the proven Tek:
Use 1 Level Teaspoon of Light Corn Syrup (Karo Syrup or the "no-name" store brand of Light Corn Syrup) per 100 ml of purified water. Mix it well to distribute the heavy syrup into the water.
You can use an empty syringe to measure out 100cc, many jars are graduated
The Karo jar is made up and Sterilized with a solid dome lid, inverted so that the seal side is up (we don't want to can it) and the band screwed on loosely. Foil Cover of course. PC time is 20 minutes.
To shoot the jar I place it in the Glovebox, remove foil, band, and lid, and shoot directly into the liquid. Inoculate at the rate of 1 cc spores per 100 cc Karo Solution. I pre-sterilize some extra jar lids wrapped in foil and I open one in the glove box and place the clean dry lid on the jar mouth with the seal side _down_. Screw the band on tight and replace the foil cap.
To Clone you open the jar and drop in (or shoot with 9er Tek) a sample of macerated tissue. I use the same basic tek when working with spore prints: open the jar, tip the print over give it a flick or a scrape... Plenty of spores drop, rehydrate, and germinate... Incubate, in a 7-10 days you are ready to shoot dozens of jars... Bulk Grain... Store the excess in the fridge, it keeps for a good long time.
Karo jars prepared in this fashion can be shaken vigorously without leaking or contam problems. Everytime I open a jar in the glovebox I always replace the dome lid with a clean sterilized one, seal side down.
It's a super Tek that gets around the problems with honey. Karo is cheap, easy, and to the point. The speed blows the doors off of Agar Culture.
I will give up some liquid culture pointers... You only need 100cc or so (I mean that's enough to shoot 100 jars already). And don't let it get thick, it reduces the refrigerated storage life (because all nutrients are consumed). All you need to grow out and shoot is a mass of mycelium about the size of a quarter in 100cc of liquid.
What you are looking for is wispy, puffy, sometimes ropy white stuff that clumps and hangs on the bottom. You should shake and/or swirl the jar on occasion to try to keep the mass broken up. Before drawing up syringes to shoot jars thrash the mycelium with the needle and break it up (it fragments very easily at this stage). The solution in the syringe need not be dense... But you should see plenty of tiny little "floaties" in there... It's all you need.
Shooting Dextrose/Karo Tek early helps in a few ways: it saves time; it prevents heavy clumping which can clog standard spore gauge needles; the residual sugar, freshly macerated tissue, and change of media gives a "hot" inoculation, showing growth in 24 hours under incubation; and the cultures appear to store better when fridged. The longer you hold them the greater the chance of Contam.
This is a good place to drop another tek tip: Make sure the jars and everything are crystal clean... Not that a bit of filth hurts the tek (because it is pc sterilized) but flecks of dirt in the clear liquid culture _look_ like contams... and you never seem to notice them until after the mycelium has started to take off... You will be swirling in front of a good light and all of the sudden there is a flake of verm or a small grain of rice flour that somehow got in there and then it seems to make all the difference, it's very noticeable... Always difficult to identify.
I was embarrased to find a hair and another fleck of filth floating in one of my Karo jars the other day. Of course they are sterile, I forgot to wipe the jars out before filling... You would be surprised at the dust and crap that gets in the jars at filling time if you are not clean... It does not hurt the tek, but motes of dust and stuff look like contams. In fact motes of cotton dust floating look very much like spores germinating at 24-36 hours. As long as the specks are not growing the culture is fine, but with an eye after a jar or two, you will see fine detail under good light.
You can also add some small chunks of clean broken glass to the jar before sterilizing, shaking the jar with the glass in it fragments the mycelia nicely and speeds up the growth.. But it makes it harder to eyeball.
Another good way to break up the mycelium is simply to draw in a syringe and squirt it back out intot the liquid. This usually disperses the myc very well and helps to break up the larger clumps. As for volume. I use 200-250cc (a pint jar about halfway). I shake every day for aeration and to prevent clumping and matting. When its done in about 12-14 days or so, at rest, the mycelium reaches about 3/4 of the way up from the bottom of the jar. This ensures that all 20-25 syringes have ample mycelium. Since the mycelium is actually cultured in the liquid, any syringe will grow out even if there are no visible mycelium clumps (as opposed to say 9ers cloning tek where if you disperse the tissue fragment in too much water, some syringes may have no mycelium fragments and will not grow anything) but the clear syringes usually don't blast off the same way as the relatively cloudy ones.
Hope you find this info helpful! Happy Shrooming!
Note: Pure Dextrose is still the best imo, and I do prefer it. The formula for Dextrose Tek is 1 Level Teaspoon Powdered Dextrose (malt works too) per 75 ml of water, or 1-1/4 Teaspoons powdered Dextrose in 100 ml water. Dextrose can be obtained from any homebrew supply or on-line at www.ebrew.com Dextrose is also known as Corn Sugar (suprise!). Karo contains some additional sugars (fructose),some salt, and some flavoring (vanilla) generally, so it is not quite as pure as powdered dextrose... But it is still far cleaner than honey IMO.
BTW on the same page of that link you will also find Gypsum and Lime
Timothy Leary (Timothyleary)
|Posted on Thursday, November 08, 2001 - 06:34 pm:||
here are some pics of Karo/corn syrup that the cloning tek calls for. This is not the "Karo" brand, it is the generic brand. The generic is cheaper, and clearer(in this case n e ways)
it can be found by the maple syrup in all gorcery stores (okay maybe not all, but i would say 95%)
|Posted on Thursday, November 08, 2001 - 10:24 pm:||
I find that with clear solutions I can hold them up to the light and see threads of mycelial growth when swirling gently... After just 24-36 hours with spores using a loup or magnifying glass and good light. Most of the spores germinate at 3-5 days. 7-10 days and it should be ready to shoot. If it takes 14 days or longer before the culture is ready to shoot something is wrong. Karo against pure Dextrose using clone mycelia ranks Karo only a few hours behind the Dextrose, and after several days I was unable to determine any difference between the two culture types.
Keep me posted guys.
Mr. B (Argonaut)
|Posted on Sunday, December 16, 2001 - 03:32 am:||
That's Kayro one week old.
|Posted on Sunday, December 16, 2001 - 03:59 am:||
|Posted on Sunday, December 16, 2001 - 04:46 am:||
That is exactly what you are "shooting" for
Crystal clear culture solution, wispy floaty bits of mycelial tissue. No chance of a contam hiding in one of these jars when lit from below or behind with good light and glassed over carefully with an eyeball. After inoculating and glassing over a few Karo jars you will get a feel for the Tek and a trained eyeball to boot. You can clearly see _near microscopic_ particles in one of these jars.
|Posted on Sunday, December 16, 2001 - 02:35 pm:||
Jose Cruz (Vato)
|Posted on Thursday, January 03, 2002 - 07:36 pm:||
Have never tried the karo tek, and am wondering how to inoc. and how many spores? Going to try w/ prints and w/ syrenge. How many cc's per 1/2 pint? Also how much of a print per jar? Amy help is greatly appreciated.....Vato
Dr. Cubesis III (Newbieshroomer)
|Posted on Friday, January 04, 2002 - 12:00 am:||
Karo tek IS THE way to go.
Fill a spore syringe with viable juice.
follow karo tek. I believe calls for 100 ml of water, i teaspoon of karo... after sterilizing jar and cooling. innoculate jar in glovebox or in stove. inject 1 cc ( or less ) of spore juice in.
place somewhere dark and cool your heels for a few days.
Voila!! follow purge's karo tek for the rest
(Message edited by admin on March 21, 2004)
An guy (Boomer)
|Posted on Friday, January 04, 2002 - 12:05 am:||
just a little side note:
I have done several jars, and am to the point where I like steaming a honey jar way better than microwaving it.
Miking it works, but, at least with my oven, it caramelizes much easier, whereas steaming it, and pcing, I presume, it comes out much better.
If you got the time, I'd steam/pc it before miking it. So far, it's been harder to screw it up that way for me. My oven don't change power levels, and I think that has something to do with it.
Just my 02$
|Posted on Friday, January 04, 2002 - 02:16 am:||
you only need a slightly visible spot of spores in your jar... a couple of light scrapes on that print or a quarter to half cc or so of liquid.. depending on concentration... finally starting to like the karo tek... i'm never giving up on my brown rice syrup though... i love the shit... my spores germinate and grow out so fast...
Jose Cruz (Vato)
|Posted on Friday, January 04, 2002 - 06:35 am:||
Thanks to all who posted.... Much thanx, And advice taken...Vato
|Posted on Friday, January 04, 2002 - 06:44 am:||
Im partial to dextrose for the sugars . It's completly clear , it makes it nice and easy to spot contamination. Not to mention use a litle extra helps speed colinization in the jars. Has worked very well for me so far .
Dr. Cubesis III (Newbieshroomer)
|Posted on Friday, January 04, 2002 - 05:16 pm:||
Brown rice syrup????
New to me, what is that?
|Posted on Friday, January 04, 2002 - 10:41 pm:||
lundberg's "sweet dreams" organic brown rice syrup, i like it personally... but can't really recommend it to anyone else as it is cloudy as all hell and pretty much a guessing game as far as contams go... you never know until you shoot... haven't done enough real "experimenting" with it to be able to give hard facts... it just "seems" to work really well
|Posted on Monday, November 11, 2002 - 07:43 pm:||
The impurities are likely adding to your success with pure sugar solutions. Mycelium needs trace elements and a nitrogen source to thrive, which are sorely lacking in carbon rich sugar solutions. I am suprised that there are any successes using pure dextrose or even relatively pure corn syrup. Also the sugar to water ratios are a bit high and can cause osmotic stress to the mycelium cells which will slow growth.
A few years ago I tested a series of dilutions of potato dextrose broth to see what the dilution was for best growth. I started with the below stock solution.
Stock solution (100%)
250ml potato water from boiled potatoes
1 tsp dextrose powder from the brew store
I then did rough serial dilutions of 50%, 25%, 12% and 6% as shown in the image. The blue things are magnetic stir bars. The jars are illuminated from the bottom to show detail.
Suprisingly the 25% dilution was the thickest and fastest growing, but even the minimal dilution of 6% had good growth. More than enough to be used for innoculation purposes. It might even be better since the thinner mycelium is easier to draw up into a syringe without clogging and doesn't form the thick mat on the surface. The 25% eventually formed mushrooms on the floating mycelial mat, but I didn't take a picture
Currently I use a teaspoon of malt per 500 ml of water in a quart jar. Its a bit thick when colonized (like jello) but its an easy measure and it works fine for my purposes. Malt, like potato water is unrefined and contains all necessary elements for healthy fungal growth. It is no accident that you don't see recipes for honey water, dextrose or karo agars.