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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: Violet]
    #24741437 - 10/27/17 04:21 PM (3 years, 3 months ago)

Exactly Violet, that's another cool thing about the method, you can narrow down the genetics so much faster than taking transfers with the scalpel, since the biopsy tissue is orders of magnitude smaller than a wedge. I love me my scalpel though, but it's a good way to start off from a ms plate. I'd like to say more about this but I'm busy right now.

Yeah Tom I'm aware the main reason a cloudy broth is not recommended is because it does not allow you to visually inspect the LC, in the beginning at least, until myc clears up the broth as it grows. You can also tell a lot by just observing how the colony grows, cloudy broth or not. I would not dare using a cloudy broth if I was to do LC's the traditional way but noccing with the biopsy is another story, it gives me so much peace of mind, and also gives me the license for making a nasty broth.


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OfflineAntimatter
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24741448 - 10/27/17 04:25 PM (3 years, 3 months ago)

Very cool. Thank you for sharing.

Question for you Josex,

Have you noticed if rice flour makes the LC clearer than say oats or rye?


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: Antimatter]
    #24741452 - 10/27/17 04:27 PM (3 years, 3 months ago)

That's the cloudiest ever :lol:, very "milky".

If you want a clear broth try good 'ol LME.


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24741547 - 10/27/17 05:09 PM (3 years, 3 months ago)

Quote:

mushboy said:
Quote:

hamloaf said:
Heck ya, Josex.  The biopsy LC method is an old technique.




I knew of the noob method where you stab a fruit and squirt the tissue into lc...  But not josex' method.

I am currently using his method.  Should have fruits within a week! Thanks josex for the sacred knowledge:stoned:

Super easy and awesome :thaaannks:




Creamcorn wasn't a noob. I do like this method better though. Obviously less of a contamination risk.


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Re: The Biopsy Method: A different way of doing LC. [Re: FooMan]
    #24741597 - 10/27/17 05:31 PM (3 years, 3 months ago)

You can also let a plate pin and do creamcorns style:cheers:


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OnlinemushboyM
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Re: The Biopsy Method: A different way of doing LC. [Re: cronicr] * 2
    #24741610 - 10/27/17 05:36 PM (3 years, 3 months ago)

noob method because it involves shooting dirty fruit tissue into karo water.

and this gem..

Quote:

Quickly wipe your needle with iso (flame if desired, I don't bother - we did just sterilize it after all!)




:facepalm3:

although it was 11yrs ago. back then i was a noob too. now i use josex method and i am the master.
:sagetrip:


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24744453 - 10/29/17 12:38 AM (3 years, 3 months ago)

Wow, just read Creamcorn's tek, just wow! haha
How in the actual fuck can someone manage to pull that off without contaminating the LC? :facepalm3:

It's almost offensive to compare it with this Ham. :ohyou:

This tek is not about taking a perfectly visible core sample either but about inoculating with literally microscopic pieces of tissue from a clean agar culture to make the procedure as safe as possible, so you can perfectly get away with no testing and dicking around.

Also, I would not do this method poking an invitro agar pin as Cron said, I'd transfer the pin to a blank first. My confidence in pp5 containers with snap-on lids has shaken quite a bit after my last mold disaster a few months back. They're far from perfect and shit can def get in them if you keep them for long till they pin.


Edited by Josex (10/29/17 01:05 AM)


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OfflineJpro54
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24744637 - 10/29/17 03:40 AM (3 years, 3 months ago)

Will have to try this. Not into LC yet but making my way there. See if an LC newbie can do this without somehow messing it up.

Good write-up Josex


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Re: The Biopsy Method: A different way of doing LC. [Re: Jpro54]
    #24745139 - 10/29/17 11:53 AM (3 years, 3 months ago)

im real excited to try this method as i havent had much luck with LCs in the past. im going to try a slightly modded method and use one of my insulin syringes just for shits and giggles, note the needle gauge is super small. im talkin 31g. if it all works out ill post some results.


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OnlinemushboyM
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Re: The Biopsy Method: A different way of doing LC. [Re: ASliceofPI] * 1
    #24745163 - 10/29/17 12:03 PM (3 years, 3 months ago)

i used a flat headed needle so i had to be careful not to take a 'core sample' as josex mentioned.

you want to have a few invisible myc fragments stick to the heated needle and nothing else.
you are not taking a 'agar sample' but a 'culture' sample.:thumbup:

back to you josex:thaaannks:


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InvisibleViolet
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24745167 - 10/29/17 12:04 PM (3 years, 3 months ago)

Quote:

Josex said:
Also, I would not do this method poking an invitro agar pin as Cron said, I'd transfer the pin to a blank first. My confidence in pp5 containers with snap-on lids has shaken quite a bit after my last mold disaster a few months back. They're far from perfect and shit can def get in them if you keep them for long till they pin.



I hope you took note of the detail I offered in my last post, about the screw-top lids even working unmodified for LC.

With the exception of fewer fitting in the cooker, I can hardly understand why a ton more people aren't using these ziploc containers for cultivation. They're like a dream come true for every utility I've found so far for Cubensis cultivation and most utilities growing in general. Essentially I find them only poorer for constant spawn production.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

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The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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Re: The Biopsy Method: A different way of doing LC. [Re: Violet]
    #24745316 - 10/29/17 01:25 PM (3 years, 3 months ago)

Quote:

Jpro54 said:
Will have to try this. Not into LC yet but making my way there. See if an LC newbie can do this without somehow messing it up.

Good write-up Josex




The inoculation procedure itself is totally noob friendly. But you need some experience with agar, that's the only reason I would not tell a beginner to try this right off the bat, at least not until they have acquired some agar skills.

Agar is also very easy, but you need some practice under your belt to be able to tell healthy growth when you see it and to spot contamination. But you could always post pics of a plate here if you aren't sure whether a culture is good to use or not, that would also speed up your learning curve. :thumbup:

Quote:

ASliceofPI said:
im real excited to try this method as i havent had much luck with LCs in the past. im going to try a slightly modded method and use one of my insulin syringes just for shits and giggles, note the needle gauge is super small. im talkin 31g. if it all works out ill post some results.




That's going to be interesting. I think a 31G needle will work just fine. Perhaps it can take longer to see growth in the liquid, maybe 5 days instead of 2 or 3, dunno. Please let us know!

Quote:

mushboy said:
i used a flat headed needle so i had to be careful not to take a 'core sample' as josex mentioned.

you want to have a few invisible myc fragments stick to the heated needle and nothing else.
you are not taking a 'agar sample' but a 'culture' sample.:thumbup:

back to you josex:thaaannks:




I told you in one of my :pm: that I hadn't tried a blunt needle which is kinda true lol. But I did a little experiment back when I started doing this. I grabbed a colonized plate I was going to trash, a 21G sharp needle, a 14G blunt needle and a magnifying glass. Then I poked the culture with the thin needle and looked through the magnifying glass but could not see anything at all, just a clean needle as mentioned in the OP. I did the same with the wide ass blunt needle, same shit, just a clean needle, I could not take a "plug" of the culture with it, but maybe you can with a thinner blunt needle, dunno.
Quote:

Violet said:

With the exception of fewer fitting in the cooker, I can hardly understand why a ton more people aren't using these ziploc containers for cultivation.




That's pretty much the only reason I use the containers I'm currently using, I can fit 120 in the pc and I currently do lots of agar. Well they're also very easy to handle in the SAB because the size is just perfect. I did not have any problems with them before until the mold problem I mentioned before. Half pint mason jars or the containers you use are def safer for agar than any type of snap-on pp5 like mini rounds (which also contaminated on me without opening them) or the ones I use right now. But the size is just a little too much I think.


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24745384 - 10/29/17 01:51 PM (3 years, 3 months ago)

Huh, I found that blunt needles worked better.  The needles are called hypodermic which means adapted for use in or administered by injection beneath the skin, they're designed NOT to cut a sample. Of course you're still touching them to your culture regardless, we throw medical use needles away after their use because they are contaminated by the contact.  But when I do a clone scrape using a needle (still often to agar) I am combining a scrape with the puncture motion, otherwise I found it just pushes through which you do not want for a fruit body clone.

The blunt tip made stab motions more successful for me in taking a cylinder, although sometimes for fruit tissue I would still end up scraping.



As for the containers.  I guess in part thanks to my clean success rate and clever little culturing steps I end up using much less agar and fewer dishes of any sort as a whole.  It's a transfer or storage medium for me, and I have gotten to running a very tight ship.  If I trimmed down to a collection of just 30 containers to turn around, I could still fairly easily operate all my gourmet culture handling and all my exotic culture hunting.

shhh....


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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Re: The Biopsy Method: A different way of doing LC. [Re: Violet]
    #24745421 - 10/29/17 02:08 PM (3 years, 3 months ago)

Just tried this with a plate of lizard king..will let you know how it worked in a few weeks


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Re: The Biopsy Method: A different way of doing LC. [Re: Violet]
    #24745438 - 10/29/17 02:13 PM (3 years, 3 months ago)

We ain't talking about taking a sample from a fruit Violet. Sure a blunt needle would be better for that.
Mushboy tried a blunt needle for this and he told me he actually saw a little fragment of agar floating around in the liquid after inoculating.

Blunt needles, sharp needles, all work for this. Sharp needles take a smaller sample from the agar culture and blunt needles seem to take a bigger sample.  It's a matter of preference really, and I like the sample to be as tiny as possible, no scraping and shit, just poking fast. When ASliceofPI posts his results with a 31G needle I may switch to those.

There are as many approaches to culturing as there are people Violet, you really don't need to shhh me or anybody :lol:
I personally would not need to do so much agar really, when all it takes to have 30 jars of clean spawn is poking a single culture with a needle. :shrug:
I do it for the fuck of it cuz I'm agar whore :shrug: and love to see the difference in growth patterns and ramifications a culture can adopt after a few generations of transfers, again, for the fuck of it.


Edited by Josex (10/29/17 02:27 PM)


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OnlinemushboyM
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #24745461 - 10/29/17 02:24 PM (3 years, 3 months ago)

i go through lots of agar myself. id need hundreds of those ziplocks but can only pc  few at a time? :obamafrown:

heres the 'core sample' floating around.

in the upper center left you can see a blurry little fuzz. if you look hard enough you can see the denser agar.


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Re: The Biopsy Method: A different way of doing LC. [Re: mushboy]
    #24745626 - 10/29/17 03:44 PM (3 years, 3 months ago)

I know, the fruit clone was brought up as an aside, but I was still using it for comparison for this as well, as to why I was surprised you commented that you figured blunt tips may not work as well.  I might not have understood perfectly if you weren't staying away from blunt needles.

I can't see any downsides to using such a tiny sample as even a 31G would take for expansion to a grow-out.

Also, I wasn't shushing you about what you're doing, that was as if I was showing you a part of a little personal secret.
What you're doing is fine, great even.  I used to boom out lots more agar plates as well, but since then I've decided just to trim it down to the smaller focus, only telling you about what I do and why I don't mind the size.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

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The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts


Edited by Violet (10/29/17 03:54 PM)


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OfflineJpro54
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex] * 1
    #24745650 - 10/29/17 03:57 PM (3 years, 3 months ago)

"The inoculation procedure itself is totally noob friendly. But you need some experience with agar, that's the only reason I would not tell a beginner to try this right off the bat, at least not until they have acquired some agar skills.

Agar is also very easy, but you need some practice under your belt to be able to tell healthy growth when you see it and to spot contamination. But you could always post pics of a plate here if you aren't sure whether a culture is good to use or not, that would also speed up your learning curve. :thumbup:"

Once my dishes are covered I'll come back and post some pics before i start cutting to make isolates. I used a good Martinique specimen to clone to agar already.


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Re: The Biopsy Method: A different way of doing LC. [Re: Jpro54]
    #24745819 - 10/29/17 05:38 PM (3 years, 3 months ago)

Giving this a shot with spelt flour tomorrow!
Dude what else are you hiding from us? Can you take a break from being a boss cultivator for a bit and do some more write ups for us please :lol:
I know there is more genius you're keeping from us


--------------------
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Our individual experiences are all part of the universe's experience of itself


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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #24746150 - 10/29/17 08:26 PM (3 years, 3 months ago)

CBK trying this!? :nowaydude:

That ain't gonna be fair man, it's gonna be like your typical CBK but on fucking steroids, watch out for this one folks!


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