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InvisibleBlueIndian
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Registered: 01/17/10
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2nd attempt with Pastyplates...need advice
    #22471861 - 11/03/15 04:16 PM (8 years, 2 months ago)

First time around I followed the tek and used a MS syringe and got no growth...nothing..not even contams. That's ok I consider it practice at least. But where I'm at now I have some jars of healthy mycelium (I hope) on oats I could take a grain and apply to the new agar plates I made. I also have some fresh fruits I could clone. Not super excited about the fruits right now as something is either wrong with the genetics or the mono tub isn't dialed in. Made the substrate pretty damn deep and wound up with some really fat ass stems and small caps...pin set was uneven and amount was pretty poor. Took some for a test run few days ago fresh and wasn't real impressed...but I've had that issue before when fresh seemed lackluster but once dried whole different story.

I followed the tek agar/water ratio today but it seemed gooey and not dissolved well. The tek amount only allowed me to make 6 plates there wasn't enough for 8. I'm trying to do some isolating and never done this before. My thoughts are to do 3 plates with colonized piece of grain and do 3 plates with clone tissue. But I need help thinking which way is best? Try both? And what is the best way to do clone tissue if I do that? I know you can use a scalpel to cut into the center but there is a tek somewhere that involves using a syringe with sterile water and jabbing it through a stem and then using that. I don't know maybe that's for making an LC.

Any help here doing it right with agar would be much appreciated..Thanks!


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OfflineDrCrumbs
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Registered: 10/25/11
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Re: 2nd attempt with Pastyplates...need advice [Re: BlueIndian]
    #22471894 - 11/03/15 04:22 PM (8 years, 2 months ago)

Is the plate underwater?

How long has it been?

Spores take time, especially based on age.


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InvisibleBlueIndian
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Re: 2nd attempt with Pastyplates...need advice [Re: DrCrumbs]
    #22471932 - 11/03/15 04:29 PM (8 years, 2 months ago)

Those old plates are long gone I gave up on them. I have 6 fresh ones I made today and have established myc to work with rather than a syringe..that's what I'm wondering is best way to go here.


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OfflineMachiavelliavore
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Re: 2nd attempt with Pastyplates...need advice [Re: BlueIndian]
    #22472815 - 11/03/15 07:14 PM (8 years, 2 months ago)

By your description it sounds like you had insufficient FAE resulting in CO2 buildup that caused deformed fruits.  Poorly formed caps and thick fuzzy stems are typical.  High abort rate is also common with intense CO2 buildup.

Fresh vs Dry:  Sometimes fruits are more than 90% water, meaning you would need to take more than 10x the dry weight to get an equivalence.  All my best trips have come from dry, even though I prefer to eat fresh.

If the agar (especially if you're using bars/sticks) is not properly distributed from plate to plate, you're gonna get some wet ones.  Perhaps you need some more heat to disolve it.  If you're confident enough in your airbox technique, you can try putting all the pasty plate mix in a jar, then pouring the empty sterilized pasty plates.  I did it, fun experience, I enjoyed it.

Don't do that syringe biopsy, it's pretty dirty.  What you want to do is tear the stipe in half, being careful not to touch the inside of one half of the stipe at all.  Use your thumb on the top of the stipe to push say the top half to the side, then wedge your thumb between the two pieces of stipe, so you can hold the mushroom and keep one piece out of the way at the same time.  Personally I prefer flamed tweezers to a blade.  Grab a fiber of mycellium that is not exposed to the outside of the mushroom, grab it, twist, and place that on a plate.  As close to the bottom of the stipe as possible is best.  Don't let your hands get above the plate or above the tissue sample #1 priority.

Definitely open up one of your plates and just see if it's wet and mushy.  Better to waste one plate than waste your time putting shit onto 6 bad plates.  It should be once floppy piece.

You can put grains on agar, or a drop of spore solution, or stipe tissue, it doesn't matter.  A single grain will most likely be a bit more genetically limited than a drop of spore solution, that's the only difference.  Clones should be your eventual goal, but wait until you have something worth cloning.

You should see some growth from MS even on wonky agar, as long as its got some starch.  IDK, perhaps a bunk syringe.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


Edited by Machiavelliavore (11/03/15 07:24 PM)


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