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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy] * 1
    #24741137 - 10/27/17 11:37 AM (6 years, 3 months ago)

this is so awsome and well written and definitely needed to be added to the shroomery.

Great contribution and great clean safe easy lc method.

This is how I will do my broths in the futur although I like pasty's ez lc tek as well.

This just seems a bit easier and less vectors.

Great idea and thanks for sharing. You F'n ROCK!:eek:


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: tombosley8]
    #24741191 - 10/27/17 12:05 PM (6 years, 3 months ago)

I may try this with pasty's ez lc as well for spreading a plate into tons of lc with barely any work/inoculant.

Seeing as mushboy got good growth on agar I should have no problem with recovery on ezlc, right?


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24741337 - 10/27/17 01:34 PM (6 years, 3 months ago)

Thanks for the addition as I think I may have been over thinking it.

The main reason for the ezlc is to make sure the inoculation process and innouculant were clean before adding the liquid.

It's just a safe check but you still have to add the water after so you could potentially contam and not know at that stage.

So now that you helped me get through it I will most likely just use the broth method for this tek.

the only other thing to the ezlc is you can use better sources of nutrients without clouding up the liquid. Your grain water lc is pretty much as good as you can get except it has the cloudiness. Again that only would matter to identify a contaminated lc.

I have had a couple instances where I wouldn't have known my lc was bacterial but could see the growth before adding the liquid. Probably bad technique or possibly the culture, it just showed the bacteria better in the ezlc than the original agar plate.

I'll probably do mostly broth and just test it on agar as you said in the op.

Again thanks so much for this and it should prove to be immensely helpful


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex] * 1
    #24750001 - 10/31/17 01:44 PM (6 years, 2 months ago)

Yeah mushboy Rockin IT!

Can't wait to try :thumbsup:


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: insanemike]
    #24768769 - 11/08/17 09:52 AM (6 years, 2 months ago)

this tek is using agar not fruits for inoculating the lc


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24768819 - 11/08/17 10:11 AM (6 years, 2 months ago)

damnit you guys are making me really want to try this.

I guess tomorrow is a good day!


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: JHOVA]
    #24768980 - 11/08/17 10:52 AM (6 years, 2 months ago)

I'd say lc is the fastest easiest way for noccing cakes.

I wouldn't risk the stipe tissue inoculation in anything but agar.


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24768995 - 11/08/17 10:58 AM (6 years, 2 months ago)

for me using a thick lc is about 4-5 days with 15 ccs poured over the top and down the sides.

I've gotten 3 days before but the growth is light untill day 4 or 5.

Either way 3 days vs 6 days isn't going to change much. just make/inoculate the cakes ahead of time so the couple extra days isn't going to be a problem.


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Edited by tombosley8 (11/08/17 11:13 AM)


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: insanemike]
    #24769219 - 11/08/17 12:51 PM (6 years, 2 months ago)

that has nothing to do with the thread. and I think the answer to that question has been answered on the shroomery countless times. Cutting brings in contams from the outer tissues into the middle where you want to take a clean cut from.

Ripping is done with gloved hands and in the sab and carefully so you don't have your hand going over or touching the clean inside.

ok enough about that.


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Edited by tombosley8 (11/08/17 12:51 PM)


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: herrenvolk]
    #24790039 - 11/17/17 05:26 PM (6 years, 2 months ago)

ok finally actually doing this and am sterilizing the lc broth and syringes with water as we speak. Really happy about this thanks again josex.

going to be cleaning some ms plates with your biopsy method as well.


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24790181 - 11/17/17 06:28 PM (6 years, 2 months ago)

what is the problem with losing a 1/4 of a ml of water?
Like how is that going to change anything?:shrug:


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24790201 - 11/17/17 06:40 PM (6 years, 2 months ago)

kool thanks for the tipz


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: bodhisatta]
    #24790557 - 11/17/17 09:52 PM (6 years, 2 months ago)

is there a reason you warm the water before adding LME?

I've found it dissolves better/more quickly into a cold liquid not hot.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24790634 - 11/17/17 10:31 PM (6 years, 2 months ago)

:shrug: Mine is opposite
just did both cold and hot and the hot had clumps that were harder to stir in. Had to use a wisk to get them stir in.

although possibly the malt I weighed for the hot had a couple clumps in dry form that caused the problem.

but I've used malt extract in lcs for a while and I've always found it easier to stir into cold. well actually I add the dry malt then pour the water over it.


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: bodhisatta]
    #24790687 - 11/17/17 11:10 PM (6 years, 2 months ago)

thanks for the info Bod I'll add the dry to the liquid next time. but I'm glad cold works for me either way. but i'll try luke warm to see the difference as well.

Definitely not trying hot again, no offense Josex but maybe it is the amount I'm making at a time or the type of malt that we are using differs somehow. just had to check one last time to know for sure I wasn't crazy.

I did a couple with the grain flour too and I'm liking them.

I rarely can notice a contam from visual so the cloudiness isn't a problem for me. I just was always scared of it for some reason.

well mainly because when I had really shitty technique, like noccing with spores, I would watch certain ones get very cloudy very quickly with nasty smelling bacteria.

haven't seen that for a long time and I trust my technique enough to not need that clear of a broth.

Thanks again buddy I think these are my new gotos!:hatsoff::thumbup:


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24801743 - 11/23/17 08:09 AM (6 years, 2 months ago)

Will do some magic with this one
ms lgt 5th xfer


Edited by tombosley8 (11/23/17 08:10 AM)


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Offlinetombosley8
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24801825 - 11/23/17 09:13 AM (6 years, 2 months ago)

yep exactly. Thanks for playing. :cultureclash:

and also going my old route with it from Li to lc broth except using your grain lc broth instead of mea...

Definitely one of my better/best looking lgt plates so I put it to a lot of good use "magic"


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Edited by tombosley8 (11/23/17 09:16 AM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24801839 - 11/23/17 09:22 AM (6 years, 2 months ago)

Thanks I needed that!


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Re: The Biopsy Method: A different way of doing LC. [Re: TheMadHatter420]
    #24801849 - 11/23/17 09:27 AM (6 years, 2 months ago)

Not seeing any growth from 4 days ago on a couple biopsy Lc inoculations. It's kinda hard to see through it, I thought I saw a small dot of floating myc but then I lost it really quick. Think I'm just overly anxious.

oh yeah haven't seen any growth on the agar plates either that were nocced with the biopsy 4 days ago.

Sure something will pop up soon/
oh yeah the wheat is much clearer than oat lc


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Edited by tombosley8 (11/23/17 09:28 AM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Fungus Mountain]
    #24848780 - 12/15/17 09:25 PM (6 years, 1 month ago)

unfiltered with metal lids rubber side down?

i'VE BEEN THINKING of doing some lcs(well I already did one) in qt jars with a good amount of airspace above the liquid and unfiltered unmodded plastic lids.

Should that be enough air for proper growth of say 400ml of lc or will it stall or not grow properly due to lack of ge?


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