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OfflineWronguy
Registered: 03/05/05
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Requesting Liquid Culture recipes for FAQ. I need as much input as possible.
    #5889987 - 07/23/06 11:29 AM (15 years, 2 months ago)

I will be adding a Liquid Culture recipe section to the FAQ and need as much help from you guys and gals as possible. Liquid cultures are becoming very popular and we need to have a detailed section covering this for the benefit of everyone.

Please post your recipe, instructions for preparation, incubating practices, agitation practices, and even a photo of the finished product would be very helpful.

I'll start this one off.

Recipe:
Using a full coffee pot of water (12 cups), I use 10 grams of dextrose and 10 grams of light malt extract for every 4 coffee cups of water. I am specifically saying "coffee cups" because a coffee cup is less than 8 oz. or a standard cup. Okay, so that equates to 30 grams of dextrose and 30 grams of light malt extract for every full pot.

Place the dex/malt into the area where you load the coffee using 2 coffee filters. Run the machine. This will yield 12 cups of liquid culture medium. Load these over 3 quart jars, cover and PC for 25 minutes at 15 PSI. Allow to cool overnight.

Inoculate each jar with 1cc of spore solution. Agitate the solution everyday with either a magnetic stirrer (stir bars loaded prior to PC'ing) or with marbles prior to PC'ing. You should have healthy colonization and be ready to inoculate within 7-10 days. Your solution should like something like this when it's ready:


South American strain

This recipe was concocted with the help of Blue Helix.

If you are posting someone else's recipe, please make sure they are in fact the recipe inventor, as we need to give credit where credit is due. Thank you.


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Invisiblecreamcorn
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Wronguy]
    #5890007 - 07/23/06 11:40 AM (15 years, 2 months ago)

2-4% (but absolutely never over 4%) honey or corn syrup (karo) by weight. easy rule of thumb, 4g of honey or karo per 100ml's of water. between the rounding and the fact that there's some water in honey/karo that lands you in safe territory just under 4% sugars.

ive got more details about how i prepare LC in my LC cloning link below...

pictures from there you're free to use:


colonized half pint (100ml), without agitation = big cloud


colonized quart (600ml), with constant mag stir agitation = lots of little clouds

both B+ strain, 4% honey in distilled water


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OfflineWronguy
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: creamcorn]
    #5890019 - 07/23/06 11:46 AM (15 years, 2 months ago)

Creamcorn, I will be posting these directly into the FAQ, so please cut and paste the pertinent information in your thread, in the order of preparation. Thank you for your input.


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OfflineYamidude
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Registered: 06/15/06
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Wronguy]
    #5890032 - 07/23/06 11:52 AM (15 years, 2 months ago)

good call I was looking for some info the other day before i started my LC project and only found limited info..


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InvisibleTippinthru
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Registered: 04/07/05
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Wronguy]
    #5890139 - 07/23/06 12:37 PM (15 years, 2 months ago)

First, modify LC jar lids with a good injection port, and gas exchange provision.

Low tek, is to drill two holes opposing each other on the lid. One covered with silicone, as an injection port, use adhesive to apply a tyvek patch to the top, and underside of the other hole. Doing so, provides a double barrier, with a lid width air space between them.

Or, any method, you can improvise.


A level tablespoon each light dry malt, and dry dextrose to a quart of boiling water in a pot. Stir in, and allow to stay at a rolling boil, for a minute, or 2. Pour half that mixture through 4 or 5 nested coffee filters, into 4 pint jars, or a quart jar. Or, all of it, into a half gallon jar.

You only want the jar, HALF FULL, to allow ample air space inside it.

If you have a stir plate, add a stir bar.

If you don't have a stir plate, add a marble or 2, as agitating devices.

Optimal sterilization, is to PC the jar, or jars 15 to 20 minutes at 15 psi, with the lids 1/4 to 1/2 turn loose.

Allow to cool inside the PC, at least to the point, pressure drops to zero, plus 15 minutes.

Open PC, and quickly tighten the lid, or lids.

(Microwave to boiling, several times, will also work.)

Once done, store in a clean place, until cooled to room temperature.

Inject 2.5 ml spore solution into each, and incubate at 74 to 80F.

Agitate, daily.


Example of a 1/2 gallon jar, with a quart LC inside it.

Light dry malt, and dry dextrose can be purchased at beer brewing supply stores.
Dry dextrose, is corn sugar.
If you cannot find those ingredients, Karo syrup will work.


Grain, wbs jars injected with LC colonize FAST.




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Perfection is attained by slow degrees; it requires the hand of time...
[


Edited by Tippinthru (07/23/06 01:12 PM)


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Offlinejcee
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Tippinthru]
    #5892163 - 07/23/06 09:55 PM (15 years, 2 months ago)

how would you suck up lc without having turn it upside down. Im about to run into this prob with my lc. I have micropore tape covering a hole for air exchange and Im afraid to turn the lc upside down for it would surely leak and ruin my lc. would tyvek leak if you flip upside down?


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InvisibleTippinthru
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: jcee]
    #5892231 - 07/23/06 10:09 PM (15 years, 2 months ago)

You have the injection port on one side of the lid, and the gas exchange filter on the other (opposing each other).

You insert the syringe into the port, then tip the jar side ways, until the needle is in the LC, then you aspirate LC out.

Do NOT turn the jar upside down.

Do NOT get the filter material WET.

Otherwise, you are in for a calamity :eek:.

If you have high end filters?
You can get them moist.
But, it is not recommended.





--------------------
Perfection is attained by slow degrees; it requires the hand of time...
[


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Offlinejcee
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Tippinthru]
    #5892260 - 07/23/06 10:19 PM (15 years, 2 months ago)

what are you using for air exchange

if I were using tyvek on what side do I put it on


Edited by jcee (07/23/06 10:21 PM)


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Invisiblefastfred
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: jcee]
    #5892278 - 07/23/06 10:26 PM (15 years, 2 months ago)

Here ya go. That's 60 of them for you! With literature citations even.

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/3077006/

I would use creamcorn's LC cloning tek as a starting point. Other important factors are having 2-4% carbon(sugar) content. Honey and Karo are around 20% water, so 5ml/100ml is about 4%.

Also look at my LC contamination FAQ in the faq submission section. That should help people spot bacterial and yeast contamination.


-FF


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Invisibleesdfsfd
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: fastfred]
    #5892348 - 07/23/06 10:46 PM (15 years, 2 months ago)

wow good work fastfred.

1 question though, why so many different recipies?
>>nevermind i just read the discussion after your initial post

jcee..i swirl my LC just a bit then tilt and insert syringe and when a mycelium clump passes by the needle i suck it up.


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Invisiblefastfred
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: esdfsfd]
    #5892464 - 07/23/06 11:19 PM (15 years, 2 months ago)

Yeah, a lot of people dissed me saying that most of them wouldn't work for fungi. That isn't true though. I think all of them listed will grow at least some sort of fungi. I've actually revised it a bit and have a new version, but it's mostly the same so it needs more work before it will be worthwhile to post it.

I just wanted it to cover all the standard ones you might see in a journal article and to make sure that everyone could see what the standard recipe was. Almost all of them are the standard FDA approved methods, so if you make one of those you will be making the standard formulation, not something that everyone likes to make a little different.


-FF


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OfflineWronguy
Registered: 03/05/05
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: fastfred]
    #5892496 - 07/23/06 11:28 PM (15 years, 2 months ago)

Great input people! Keep 'em coming folks.


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Invisibleliamtheloser
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Wronguy]
    #5892698 - 07/24/06 12:45 AM (15 years, 2 months ago)

Here's a tech I made up... works very well too.

Every time you use up a spore syringe, you can make an LC culture out of it... Basically PC a jar of karo water, like you would for making a regular lc, but when it cools, suck the karo water into the syringe until it's full, then set it in a drawer somewhere... a week later it'll have enough myc for another few jars.  I realize this is a white trash LC tek, but hey... whatever.



basically it works because a lot of spores stick to the rubber part of the plunger, and when you introduce the karowater, they grow...

Anyways, if someone wants to type that out all professional and shit like all the other teks are, go ahead :laugh:


Also, if you want to stretch it even further, you can inject the syringe full of myc into a newly prepared karo-water jar and it will continue to colonize that.

anyways, love the addition of new teks.  I eat that shit up


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OfflineTheHulk
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: Wronguy]
    #5892700 - 07/24/06 12:45 AM (15 years, 2 months ago)

1. Pour 100ml of apple juice and 300ml of water into a mason jar.
(don't forget to use tyvek over the mouth of your jar..for protection, and don't forget your marble or metal object to agitate your mycelium)
2. Pressure cook your jar at 10 PSI for 10 Minuites
3. Allow the jar to cool for a few hours
4. Open your jar, scrape in spores from print or inject spores with syringe
5. Close cap on jar tightly, put away in a warm,dark area, use a rubberband and wrap a coffee filter over the cap of your jar to keep away contams.
6. Wait for growth, hopefully it will be white, cloudy growth.
7. Make sure that atleast once a day you stir the contents of your jar
8. Don't let it grow too thick, it's done when about 80% of the liquid is made up of the white cloudy stuff.

Using this method I see growth in 24 hours, using a magnetic stirrer helps speed up further growing.

For my magnetic stirrer I use a homemade stirrer made from a computer fan and a magnet from a hard drive. Search the shroomery for details on how to make your own magnetic stirrer. You probably have all the parts needed to make a stirrer laying around in your home.


--------------------
long live shroomery, much love :smile:


Edited by Mr_Big (07/24/06 01:05 AM)


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Invisibleliamtheloser
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: TheHulk]
    #5892712 - 07/24/06 12:51 AM (15 years, 2 months ago)

40 minutes of pcing? that's more than you need for a pf cake... how about 10 minutes


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OfflineTheHulk
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: liamtheloser]
    #5892729 - 07/24/06 01:02 AM (15 years, 2 months ago)

I'm a total noob, pwn me plz, I pressure cooked for 40 minuites just to be safe. I will not spend so much time pressure cooking my LC from now on, thanks for the info. I'm new to cultivating and found this method very easy, and working fine everytime. So I decided it would be a good idea to post it, if it worked for one newb it should work for others.


--------------------
long live shroomery, much love :smile:


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Invisibleliamtheloser
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: TheHulk]
    #5892746 - 07/24/06 01:09 AM (15 years, 2 months ago)

Yeah, but 40 minutes will carmelize the fructose in your apple juice.

10 minutes is good for sterlizing liquid.

15 minutes to play it safe

40 minutes is too much.

I don't consider this pwning you, I consider it informing someone with great intentions... you should feel honored because I've pwned greater men.


--------------------


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Invisiblecreamcorn
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: liamtheloser]
    #5892774 - 07/24/06 01:23 AM (15 years, 2 months ago)

i think he was referring to your tag line, not requesting a personal pwning.  :cool:


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Invisibleliamtheloser
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: creamcorn]
    #5892781 - 07/24/06 01:25 AM (15 years, 2 months ago)

no, he was requesting it... it's hard to resist getting pwned by such a pwntenating master.

However, I resisted, and I feel like a better person.

Watch out creamypants, I'll pwn you next :p


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OfflineYamidude
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Re: Requesting Liquid Culture recipes for FAQ. I need as much input as possible. [Re: liamtheloser]
    #5892796 - 07/24/06 01:28 AM (15 years, 2 months ago)

penis shits.. Back to the LC..


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