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InvisiblebodhisattaM
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Re: The Biopsy Method: A different way of doing LC. [Re: van hatton]
    #24752923 - 11/01/17 08:36 PM (3 years, 4 months ago)

ended up doing this the other day. Felt like a couple jars didn't get anything, a couple jars definitely did. I would have went back to re-inoculate the ones I was concerned about but I want to see if it will just work anyway.

I also made one LC with a small 1cm square wedge.

I use metal lids rubber side down, I never have really hard a problem breaking that seal and putting wedges in jars.


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Re: The Biopsy Method: A different way of doing LC. [Re: bodhisatta]
    #24752931 - 11/01/17 08:43 PM (3 years, 4 months ago)

Strange Bod, it hasn't failed me a single time yet, how long since inoculation?


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24753022 - 11/01/17 09:40 PM (3 years, 4 months ago)

Here's my 10/29 - 2 aerial myc twisted pair threads:

11/1



Donor plate was T3 from an oat that pinned invitro


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InvisiblebodhisattaM
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24753029 - 11/01/17 09:43 PM (3 years, 4 months ago)

Quote:

Josex said:
Strange Bod, it hasn't failed me a single time yet, how long since inoculation?



Not even 24
Hence time will tell. I just feel like nothing got shot in. Something probably did


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Re: The Biopsy Method: A different way of doing LC. [Re: bodhisatta]
    #24753043 - 11/01/17 09:49 PM (3 years, 4 months ago)

That's just part of the game :lol:

Quote:

Now I'm going to talk about your feelings.
After the poke, you may feel skeptical, as if you just did nothing, cuz no way in hell this is going to work...




Even I still feel the same every time I do it, best thing is forget about the jars and come see them 4 days later.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24753054 - 11/01/17 09:55 PM (3 years, 4 months ago)

I think I would feel more comfortable with a blunt needle tip. But youre right I'm probably just trippin :lol:


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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #24753088 - 11/01/17 10:06 PM (3 years, 4 months ago)

If you use a wide needle or blunt needle, you're missing out on the magic man. It's also tougher to fuck up with a thin needle, and what about growing pounds of shrooms from a piece of literally microscopic tissue.
:magicfingers: 

Well shit, if those 31G needles for diabetes came with standard hubs for standard syringes I'd have ordered them by now.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24754055 - 11/02/17 11:45 AM (3 years, 4 months ago)

So genius man, I love it :toast:

I've been sticking to blender Li because of ease but this looks very intriguing


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24754320 - 11/02/17 01:29 PM (3 years, 4 months ago)

Josex
This could be ground breaking..
It's genius.....

I just did a turkey tail and B+ (LC), With a 21g Needle...
Wow...! Perfect clean micro punch holes in the culture...

I now thinking this could be a way to do plate transfers also...

I have a method where I tap the top of the plunger on the syringe just to get a couple of drops into my plates, without flooding or over doing it..

This could be a perfect way to transfer cultures plate to plate....


Edited by micelio (11/02/17 04:04 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: micelio]
    #24754370 - 11/02/17 01:56 PM (3 years, 4 months ago)

Someone else has posted this method as I read about it last winter, that is unless you mentioned it and didn't do a tek until recently. I did it, but alas my culture was dirty and I was to knew to agar to catch that there was still bacteria hiding in the mycelium. I do feel like this would be a good way with a clean culture.


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Re: The Biopsy Method: A different way of doing LC. [Re: TheMadHatter420]
    #24755086 - 11/02/17 07:31 PM (3 years, 3 months ago)

Yeah Mad, I had it sorta hidden in my journal, sloppily written.

Anyway, you can test a culture before using it. I posted a way at the end of the OP, take a look. Good luck man.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24755100 - 11/02/17 07:37 PM (3 years, 3 months ago)

I think I will give this biopsy to LC method a try again once I am happy with a culture.


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Re: The Biopsy Method: A different way of doing LC. [Re: micelio]
    #24755102 - 11/02/17 07:38 PM (3 years, 3 months ago)

Quote:

micelio said:
Josex
This could be ground breaking..
It's genius.....

I just did a turkey tail and B+ (LC), With a 21g Needle...
Wow...! Perfect clean micro punch holes in the culture...

I now thinking this could be a way to do plate transfers also...

I have a method where I tap the top of the plunger on the syringe just to get a couple of drops into my plates, without flooding or over doing it..

This could be a perfect way to transfer cultures plate to plate....




It is, you could get an isolate ridicously fast. A combination of using the poke and the scalpel is killer.

Two transfers with the needle followed by 2 transfers with the scalpel is the way to go about this imo, rinse and repeat.

Quote:

The picture shows growth on agar from a biopsy sample.
As you can see, growth looks disorganized, like a streaked plate from spores would. Don't be discouraged by this, it's perfectly normal and fine.

A biopsy sample taken from a rhizomorphic culture will also show tomentose and disorganized growth, but the potential for rhizo growth is still there, nothing has been lost.
Such a tiny piece of tissue needs time to regroup and establish a strong network.
A couple of transfers from this plate using a scalpel will get things back to normal, the culture will show rhizos again and you will also notice a substantial change in appearance from having narrowed down the genetics considerably.
Again, I don't have pictures at the moment to back this up, but will do soon. My apologies.




Edited by Josex (11/02/17 08:01 PM)


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Invisiblemicelio
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24755457 - 11/02/17 09:43 PM (3 years, 3 months ago)

Quote:

Josex said:
Quote:

micelio said:
Josex
This could be ground breaking..
It's genius.....

I just did a turkey tail and B+ (LC), With a 21g Needle...
Wow...! Perfect clean micro punch holes in the culture...

I now thinking this could be a way to do plate transfers also...

I have a method where I tap the top of the plunger on the syringe just to get a couple of drops into my plates, without flooding or over doing it..

This could be a perfect way to transfer cultures plate to plate....




It is, you could get an isolate ridicously fast. A combination of using the poke and the scalpel is killer.

Two transfers with the needle followed by 2 transfers with the scalpel is the way to go about this imo, rinse and repeat.

Quote:

The picture shows growth on agar from a biopsy sample.
As you can see, growth looks disorganized, like a streaked plate from spores would. Don't be discouraged by this, it's perfectly normal and fine.

A biopsy sample taken from a rhizomorphic culture will also show tomentose and disorganized growth, but the potential for rhizo growth is still there, nothing has been lost.
Such a tiny piece of tissue needs time to regroup and establish a strong network.
A couple of transfers from this plate using a scalpel will get things back to normal, the culture will show rhizos again and you will also notice a substantial change in appearance from having narrowed down the genetics considerably.
Again, I don't have pictures at the moment to back this up, but will do soon. My apologies.







:thumbup:

Much appreciated...
People like you and all the others are the reason I can't stay away from this site.....

You did the most wonderful job explaining your system.. So easy to understand...
Most of the time I see mountains of text, I go...: here we go again...  Boring...

I'll be looking for more text from you...  Thanks.....


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Re: The Biopsy Method: A different way of doing LC. [Re: micelio]
    #24755515 - 11/02/17 10:04 PM (3 years, 3 months ago)

:cheers:


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OnlinemushboyM
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24756540 - 11/03/17 11:36 AM (3 years, 3 months ago)

once these knots/pins grow a but ill post some more pics.


lucy/aa+ pins never show up too well on my camera. just looks like white blurs:tongue:

id also like to note..

due to my shaky fucking hands, MOST if not all of my contams happen during inoculation.
either from a bad culture drop, or i bang something.. usually 1 jar out of 10 gets an
out break of green on the surface of the grains. but with pouring?? not a single jar
was visibly contaminated. although 1 jars recovery took 4days...:cool: meh. point is?

this method is awesome for anyone who has a hard time with being clean(inoculation wise). cause im sloppy as fuck sometimes.


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24756586 - 11/03/17 12:04 PM (3 years, 3 months ago)

:creepylurker:

That's alright man, 4 days recovery is pretty normal, usually it takes 3 for me. After 24-48 hours you can maybe observe some growth on the surface of the jar from pieces of myc that landed on the surface. But the real recovery starts when the liquid at the bottom of the jar is slowly taken up by myc, when the bottom is colonized (4 days from inoculation, give or take), I like to give the jar the first shake. :thumbup:


Edit: I think you were referring to recovery as a test right? Like when you shake the jar at 90% colonization or whatever...
Jeez I need to pay more attention when I read.


Edited by Josex (11/03/17 12:16 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24756615 - 11/03/17 12:18 PM (3 years, 3 months ago)

haha. ya dude. the 100% test. 4days and its usually in the garbage. but i spawned it anyway:shrug:

:awegroove::popcorn:


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24757056 - 11/03/17 03:40 PM (3 years, 3 months ago)

What kind of myc density are we looking for to use an LC as an inoculant ?

Presume it scales on the amount of grain used.  Mycobag, quart, pint.

Thxx

Day 6 of two LCs PE and Pan Cyan

Pan has really taken off. Started with a dime sized transfer.

PE was a T3 half tiger drop which has gone pretty crazy in the past couple days.


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Re: The Biopsy Method: A different way of doing LC. [Re: Snazz]
    #24757061 - 11/03/17 03:41 PM (3 years, 3 months ago)

I see some recovery in most jars. I made a pretty sediment heavy LC this time tho. Pressure cook beer wort for 15m and it will be full of precipitated crud.


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