How to make a mushroom LC

For this, I am of the thinking that SIMPLE IS BEST. My liquid culture containers are simple jars, with a regular old metal lid and ring, with a single hole punched at the top. The hole should be big enough to accomodate your needle, with a little room to spare so that a vaccum is not created later when aspirating solution. The hole is covered with a piece of micropore tape, and then covered again either with a coffee filter and rubber band, or a piece of aluminum foil loosely crumpled, to act as a dust cover.

Recipe: Recipe is the same as any liquid culture; 4% media by weight. You may use any of the standard LC media, honey, caro, dex/malt will all work. Water quality is important to LC. I find that distilled works best, and gives the clearest solution. You may also use bottled spring water, or tap water filtered with a Brita pitcher or the like. (Your mileage may vary depending on the quality of your locality's tap water. In my area, I avoid it.) Honey quality also varies widely - I've found this particular variety works fantastic for me, and doesn't make any sediment in my LC's even without filtering. You may need to experiment with brands and types of honey to find one that works. For a sure thing, you can definitely use karo, and if you're industrious and willing to hunt down dextrose/malt, by all means use that. The honey I've got seems to outperform karo, so it works for me.

When cloning, I generally start with a small half pint container. You'll see I've measured exactly 4 grams of honey. Water weighs 1 gram per 1ml, so I'll fill it to the 100ml point. I nuke this in the microwave for about 30 seconds, just to warm the water, allowing the honey to dissolve easier, and I stir it up. Obviously, when making larger LC's, just do the math to stay at that 4% mark.

This will be my "master" culture. From this culture, small samples are taken to start larger liquid cultures in quart jars that are my "working" culture that I use for inoculation. The remainder of the master culture can be refrigerated, where it will keep for several months. When I run out of working culture, it only takes a few drops of the master to start a whole quart of working culture all over again. Its important to take from the master each time, rather than go from working culture to working culture, so that each of your working cultures are "second generation."

In my opinion there's only two acceptable ways to sterilize LC for consistent results: steaming, and pressure cooking. This gives those an option who don't own a PC. Microwaving works for some, but I don't trust it. If it works for you, fantastic, go with it.

In both cases, wrap the top of your jar in tinfoil as usual, and take a syringe about half full of water, and also wrap it in tinfoil. We'll sterilize them both at the same time.

Steam sterilization:
Place a cloth at the bottom of a pot with a tight fitting lid. Use an inch or two of water. Place your jar and syringe inside. Bring to a slow rolling boil. Once a boil has been achieved, start your timer for 30 minutes. At the end of 30 minutes, turn off the heat, walk away and don't even think about it for a few hours. Let it cool.

PC sterilization:

Use your PC as per its instructions with something to keep the items off the bottom of the pot (like a cloth, or a rack if you got one.) Slowly bring up to pressure. 10psi is plenty if that's all you got. 15psi is fine too. Do not exceed 15psi. Once at desired pressure, set your timer for 15 minutes. DO NOT exceed 15 minutes. When the time is up, cut your heat and walk away until its cooled.

Find a nice looking fruit! We want to clone desirable qualities. Many have different ideas as to which fruit to clone; ie. choosing the first one up might give you a speedy isolate to work with later, choosing a big one might give you a genetic predisposition towards large fruits; and so on. I believe that there's just so much variation in growing methods, even though we are taking a clone, we have no idea how closely its going to resemble its "parent" when grown out later - so I take a nice looking fruit and hope for the best. You'll find out in time if it was a good choice or not when you get your first crop of clones in, and can assess how long they took to grow, the yields you got, and of course, the potency after sampling some. If it didn't turn out as good as you were hoping, don't fret, you can try again from a different specimen from a different multispore grow. If it did turn out for you, treat that master culture like gold!

Here's where you'll want to practice your sterile procedures. No need to go overboard, all I do is ensure there's no drafts, clean my work surface, wash my hands with antibacterial soap, and hit them a second time with some hand sanitizer. If spraying Oust into the air and washing everything with bleach or whatever makes you happy, knock yourself out.

Take your fruit, and clean the stem thoroughly with an isopropyl soaked cotton swab. Clean it all the way around. Do not touch any areas with your hand near the site of the biopsy. Quickly wipe your needle with iso (flame if desired, I don't bother - we did just sterilize it after all!) and stab it straight through the stem to the other side.

Remove the foil from your prepared LC jar. Using a dry cotton swab, clean up any condensation/moisture on the lid. Then using an alcohol soaked one, clean the lid again. Give the needle another wipe, insert into hole, and push the plunger. The water in the syringe will force the cross section of the stem out into your LC. Quickly cover the hole with a piece of micropore tape (if your lid is still wet with alcohol, that's fine, the tape will stick just fine once it dries) and cover with a dust cover of your choice.

7-10 days later you should have a nice looking cloud of mycelium.

As they do on the cooking show.... "here's one we've prepared earlier!" When doing all the above steps, you'll want to work quickly to avoid contamination. I obviously stopped to photograph everything so who knows if we got a clean sample here.

Here's a "master" culture, about 10 days colonizing. You'll notice some is already missing, as I've used some of the solution to inoculate larger LCs.

Here's a working culture, this is 5 days old. It was started with about 4cc of solution from a master, and then constantly agitated on a magnetic stir plate. Use it (or make it) if you got it, but its not necessary. They'll colonize great without, it just takes a little longer. LC to LC is FAST no matter what.

-Follow the measurements carefully. You'll get no growth or poor growth if you use too much media. When in doubt, use a little less than 4%.

-If your myc cloud is thick and can't be sucked up easily, suck up what you can, and squirt it back down into the cloud. It will break right up.

-To aspirate from my single-hole jars, I just tilt them on their sides, being very careful not to spill any out the hole. The needle reaches in just fine. I gently swirl it around and capture as much mycelium in the syringe as possible. The thicker the better!

-People often ask, "when is my LC done?" Well, its done when you want to use it! If there's enough mycelium floating around that you can capture a bunch in your syringe, go for it. The rest will continue to grow until the nutrients have run out. When it doesn't seem to be growing any more, you can refrigerate it, and it lasts many months. (6 or more.)

-When I aspirate, I take an alcohol soaked cotton swab, and swab right over the piece of micropore tape. I then stick the needle right through the tape, and fill up my syringe. I quickly cap the syringe, and quickly replace the soggy piece of micropore tape with a fresh piece. You can certainly use any number of the fancy LC container teks out there, but it doesn't get much simpler than a hole with a piece of tape on it kids.

-LABEL EVERYTHING. Keep track of dates, strains, generation number, anything you feel relevant.

-A perfectly prepared karo or honey LC should be nearly crystal clear. Dex/malt will produce a yellowish solution. If your karo or honey solution turns yellow, you may have overcooked. Did you follow my sterilization directions to the T? Either way, a little overcooked will still work, its not the end of the world. Likewise, sediment from honey, or from malt/dex will not harm a thing - it just makes spotting possible contamination a little more difficult. Some take some pretty extreme measures to filter their solutions before sterilizing to get the sediments out, I'll leave it up to you.

-Unless you want cracked jars, heed my advice and allow them to cool gradually and naturally.