How do you clone mycelium when using the BRF tek?

[p88prw]

could someone please list each step that needs to be done when cloning mycelium when using the BRF tek method please ( in simple terms, as i am new to mycology).
thanks
p88prw

[Hippie3]

easiest way = take tissue sample from inside a shroom
cf. http://archives.mycotopia.net/discus...tml?1073960464

[zodd]

this how i do it

Working as sterile as possible get your cooked brf jars, your scalpel, your mushroom that you want to clone and an alcohol lamp all in one place

a bit of rubbing alcohol wont go a miss either

First off if you got rubbing alcohol rub the outside rim of the jar lid down with alcohol. Then loosen the lid a bit so you can get quick easy access when it comes to transferring the mushroom sample later.

Next flame your scalpel blade till its glowing red

Now you need to tear the mushroom in half strait down the middle. its important that you tear as opposed to cut as cutting will smear any spoors and contaminates that may be sitting on the surface all the way down the mushroom. Where as with tearing your only touching the out side stem

Next grab your scalpel and give it a second flame till glowing. now take your sample from the inner most central part of the mushroom a scraping action is best/quickest. Next you want to transfer your sample over to your brf jar. you will see now why it was important that you loosen the jar lid at the beginning.


This may take a few goes to get your sterile procedure down, but after a few it becomes second nature

[TVCasualty]

I pick an unopened mushroom from a fat cluster and cut off the cap, peel the stem like a banana, spear it with a scalpel, dip it into hydrogen peroxide (I just use a little cup of peroxide straight from the bottle), then drop it into a blender jar (it's a version of the slurry tek but you blend a stem instead of a cake). After blending it, I draw up the myc water into a syringe with the largest-bore needle I can find (minimizes clogs) then shoot it into prepared BRF jars. When the cake colonizes, I use the slurry tek to expand it to as many quarts as needed. Takes about 15-18 days to go from peeling the stem to having as many quarts as you need. Or you can stick with PF Tek style jars.

BTW: The straight 3% peroxide I dip the stem into is very strong and if you don't drop the stem into the blender (which contains about 250 mL of sterile water) right away it will likely kill the mycelium.

And be choosy when you pick the one you want to clone; I'd clone one from a dense cluster of medium-size mushrooms before I'd clone the biggest solitary specimen, for example.

[BuckarooBanzai]

I do this:

http://forums.mycotopia.net/cloning/...ek-merged.html (Buckaroo's Cloning Tek...{merged})

[Shromadon]

Well, for cloning myc, I sterilize some syringes with the jars I'll be injecting said myc into. Once its cooled down, get a store-bought bottle of water, open it up, suck water into syringe, inject into jars, swirl around 5-10 seconds, and then inject into an LC with airport. I inject the entire syringe in there just to make sure pieces of myc made it.

Don't shake the jars, just swirl, keeping the water on the bottom half. It takes practice to get the rythm down, but I only failed the first 2 times, and that was probally cuz I did not make the LC right, plus I wasn't using airports those times. You are going to want to do this in a well lit area also, cuz it can be a pain tilting the jars and getting all the liquid sucked up without it rushing into the verm layer.

I would sterilize multiple syringes, so you can use an entire one for the LC, and the rest for the jars you sterilized. When those jars start to colonize, it will look all wispy like cobweb, and won't really be white, it will be very very faded, but once fully colonizes the jars, the myc network beefs up and it gets bright white. But that is only after most, if not all of the jar is colonized.