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InvisibleComebackKid
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Re: The Biopsy Method: A different way of doing LC. [Re: herrenvolk]
    #24787939 - 11/16/17 09:15 PM (4 years, 19 days ago)

When it comes to LC... Bad LC is usually a good first guess :lol:


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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #24787974 - 11/16/17 09:28 PM (4 years, 19 days ago)

I just did some LC to jars with my really bootleg polyfil lids. and shook the fuck out of them after inoculation. we'll see.


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Re: The Biopsy Method: A different way of doing LC. [Re: herrenvolk]
    #24788069 - 11/16/17 10:06 PM (4 years, 19 days ago)

Quote:

herrenvolk said:
Quote:

bodhisatta said:
there's probably some other problem going on if a week later your jars are only just getting going. they should be done or nearly done after a week.




Yeah, like I mentioned in the other thread, I believe it was due to overly dry grains. It could, of course, also just be a bad LC. But the thing to take away from it is that although all the jars were pretty shitty, the ones not shaken were uniformly significantly better than the ones that had been shaken.




For me a bad LC is a contaminated LC, and it doesn't sound like yours was contaminated or you would've noticed by now. Your issue could be a weak culture, simple as that, and the fact that you only used 5ml per jar, which can be enough if the culture is vigorous.
The grains should look plump but dry on the outside and you should be able to move them like loose gravel, as you explained in the other thread.
Another cause could be your grains being bacterial as hell and myc not wanting to leap off.

Next time pour that bitch, gay things like syringes bring gay results, unless you're bod.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24788089 - 11/16/17 10:17 PM (4 years, 19 days ago)

Just did this with 1g LME to 500ml water :rockon: could barely see the tiny piece of myc missing from the plate after the poke but i know i got some


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Re: The Biopsy Method: A different way of doing LC. [Re: Germs]
    #24788109 - 11/16/17 10:26 PM (4 years, 19 days ago)

I think a lot of people accidentally aspirate outside air into their syringes sucking up thick LC it can leak at the connection between the syringe and needle hub.


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Re: The Biopsy Method: A different way of doing LC. [Re: Germs]
    #24788113 - 11/16/17 10:28 PM (4 years, 19 days ago)

What gauge did you use Germs? You guys need to see the stuff uh? :lol: That's not magic at all, just saying...


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24788126 - 11/16/17 10:38 PM (4 years, 19 days ago)

18, one I had laying around from a spore syringe I bought. Only shot like .5cc of water into the jar


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Re: The Biopsy Method: A different way of doing LC. [Re: Germs]
    #24788155 - 11/16/17 10:49 PM (4 years, 19 days ago)

Try 14 gauge.  14 has a much wider opening than 18.


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Re: The Biopsy Method: A different way of doing LC. [Re: hamloaf]
    #24788173 - 11/16/17 10:56 PM (4 years, 19 days ago)

Yeah, try a straw, why wouldn't you.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex] * 1
    #24788175 - 11/16/17 10:57 PM (4 years, 19 days ago)

Fuck that. Tiger drop


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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #24788178 - 11/16/17 10:58 PM (4 years, 19 days ago)

:lmafo:


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24788209 - 11/16/17 11:12 PM (4 years, 19 days ago)

If you use a wide gauage with the intention of taking an actual agar sample you're missing the point I think.

Poke with a thin needle, fast and clean and only once. I'm not even sure if anybody in this thread has done that yet. Why would you do that? No spoiler, try it and you'll see why.

Some microscopic fragments of myc will rub off on the needle and that's all you need to inoculate the broth with. In a liquid medium it works 100% of the times, even though you are going to feel like you just did nothing.
Using the same procedure to inoculate agar or pf tek pucks won't be successful 100% of the times because sometimes it will be impossible for such a tiny sample to get a hold on the agar or sub, but I'd say it works 85% of the times.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24788229 - 11/16/17 11:22 PM (4 years, 19 days ago)

No, I'm talking about for the aspiration of the liquid culture.


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Re: The Biopsy Method: A different way of doing LC. [Re: hamloaf]
    #24788256 - 11/16/17 11:36 PM (4 years, 19 days ago)

Yeah I have lots but have not used them yet, those won't clog no matter what I guess.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24788265 - 11/16/17 11:40 PM (4 years, 19 days ago)

Quote:

Josex said:
If you use a wide gauage with the intention of taking an actual agar sample you're missing the point I think.

Poke with a thin needle, fast and clean and only once. I'm not even sure if anybody in this thread has done that yet.



I did just that


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Re: The Biopsy Method: A different way of doing LC. [Re: Germs]
    #24788491 - 11/17/17 03:46 AM (4 years, 19 days ago)

12 gauge needles.


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Re: The Biopsy Method: A different way of doing LC. [Re: hamloaf]
    #24788636 - 11/17/17 07:51 AM (4 years, 19 days ago)

I don't know if this was covered, but a blunt-tipped dispensing needle makes a clean, tiny puck of colonized agar to transfer to a LC.


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Re: The Biopsy Method: A different way of doing LC. [Re: Morelman]
    #24788802 - 11/17/17 09:43 AM (4 years, 19 days ago)

I gotcha covered bro.  :douchewink:

and am i the only one who poured the LC??(following the tek)


Edited by mushboy (11/17/17 11:03 AM)


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24789120 - 11/17/17 12:20 PM (4 years, 19 days ago)

im calling this clone the 'lazy little bitch clone'


cause it takes forever to pin and last 2 tubs have been the same short n dense but spotty pinning performance.
but the yields owned me hard. and a 2g dose had my friend thoroughly fucked for awhile:stoned: ill be trying soon myself, i had work that morning:sad:

based off the other totes, this mono should be pinning in like 2 fuckin weeks.:shrug:


all open air and first time LC. i feel the fungus:thaaannks:


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Re: The Biopsy Method: A different way of doing LC. [Re: Violet]
    #24789266 - 11/17/17 01:24 PM (4 years, 18 days ago)

Quote:

Violet said:
That's perfectly reassuring. I'm gonna do syrige agar pokes for my next round of grain petris culture hunting.



Was going to reply to this at the time but couldn't and just forgot. I don't know if it will be easy for the bio to take a hold of a grain kernel, it might never grow anything. The poke fails sometimes if you shoot it on pf-tek pucks or agar and I think it would be even more difficult with grains. Try it and lets us know, but maybe you'll need a wide gauge for that.

I'm currently doing a side project using CVG+BRF pucks on little glass jars, something similar to your grain petris culturing, and only using the poke.
Started spores of KSSS, LGT and Mexi cubes, now they're two transfers away from the streaked plates (using the poke) but still on agar. Today Imma transfer them to some of these pucks (poking again), by the fifth gen of plates I'm gonna let them pin out and take clones to test. The genetics should be quite slimmed down by them and I prefer a clone taken from a culture with less of a genetic hodge-podge.

These are PE cultures that looked a litte off on agar but are perking the fuck up on this sub, pics are bad but growth is nice and rhizo, they look very happy. The poke was used to inoculate.


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