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OfflineThreads2theSoul
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27319746 - 05/23/21 11:28 AM (4 months, 27 days ago)

Thanks Josex, it all makes sense to me. I'll forge ahead to find out what works best for me, my limitations and needs, etc. The worst thing that can happen is failure and that's no big loss in this case, a few jars and supplies.


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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27319752 - 05/23/21 11:38 AM (4 months, 27 days ago)

Thank you, have fun!

Oh and yes your plan is sound in my book, except for the unmodified lids (only my opinion lol). It's not that they won't work, they will, but it's something that imo has the potential to create problems sooner or later if you do it enough.

About the last part you said, I wouldn't be cloning a clone indefinitely as you suggested. Keep the original clone culture and always go from there.

Sometimes cloning a clone can go south and you may end up with worse genetics than the original. Also, by cloning a clone over subsequent grows you'd be subjecting the colony to a lot of expansion which may weaken the culture over time.

And yeah starting from spores resets everything, doesn't matter if said spores came from an aged culture.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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OfflineThreads2theSoul
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27319767 - 05/23/21 12:04 PM (4 months, 27 days ago)

To be honest, I've been fearful of and don't have the supplies to save using a slant which from what I've gathered is the best way to save a clone. I've been reading up on LC and maybe using a thickly poured agar plate, and am going to try Fahtster's drying tek someday since I have all those supplies. Lol, all the while hoping in my hesitancy I don't loose the clone! How do you do it? Do you have a favorite tek?


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Offlinekarri0n
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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27319799 - 05/23/21 12:45 PM (4 months, 27 days ago)



Ganoderma Tsugae from a wild clone I took to agar.

Poked on 5/14 with 18ga needle, inoculated through a SHIP, and first visible signs of growth were there on day 7.


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Micropore tape? More like myco-poor tape, right? HAR HAR


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OfflineThreads2theSoul
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Re: The Biopsy Method: A different way of doing LC. [Re: karri0n]
    #27321179 - 05/24/21 03:01 PM (4 months, 26 days ago)

Would a prefilled sterile saline syringe work as opposed to sterilizing distilled or tap water?
  • Preservative free
  • Luer-lok tip
  • Sterile .9% sodium chloride

5ml 30 for $16.95
10ml 30 for $18.95

https://www.healthykin.com/p-4109-bd-posiflush-pre-filled-saline-syringe.aspx?msclkid=cecd80af730315a95201e0049fe84c08


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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27321214 - 05/24/21 03:35 PM (4 months, 26 days ago)

I wouldn't. You can get packs of 100 units of empty luer lock syringes for really cheap online, like 3 or 4 times cheaper than that.

Syringes are reusable, some of mine are years old and they're still good after having gone through a lot of cycles.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


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OfflineThreads2theSoul
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27321315 - 05/24/21 04:37 PM (4 months, 26 days ago)

Shoot Josex, I don't think I'll ever need that many syringes, 30 seemed like it would last me a long time and since they're reusable I'd have a head start on the Poke with them already filled with sterile saline solution without having to stress over sterile technique (I'll be doing that with agar and G2Gs). I figured the 5ml would be sufficient for what I'd ever need and once used, I'll start filling with water and sterilizing them for reuse. In the meantime I'd buy myself a little bit of peace of mind...  that is if the saline solution won't harm the Poke process?


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OfflineA.k.a
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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27321345 - 05/24/21 05:07 PM (4 months, 26 days ago)

There’s no sterile technique for it really, just fill the syringe up, wrap it in foil and throw it in the instant pot for 30 mins.


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OfflineSpaceBabyS
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27328530 - 05/29/21 11:55 PM (4 months, 20 days ago)

I have 6 minutes to go for PCing the syringes. B+ clone JoPo tomorrow. Stoked to poke. heheh

Finally have my PC totally dialed in. :rockon:

More later.


--------------------
SpaceBaby
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OfflineSpaceBabyS
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Re: The Biopsy Method: A different way of doing LC. [Re: SpaceBaby]
    #27329233 - 05/30/21 03:16 PM (4 months, 20 days ago)

Hey, Josex, I was thinking, dangerous, I know. I couldn’t find friigin PP5 shaker bottles anywhere. But I did find autoclavable Nalgene wash bottles, 500 ml.

I was thinking of prepping the LC in them, and doing fingers crossed noccing the Jpoke into these in the SAB.
I thought that if the original hole from the tube is too small, I can mod it with a boxcutter. :confused:

I am going to try to nocc jars with the LC while not releasing “the squeeze” so as not to draw [even SAB] air back into the bottle.

These:






Also could not resist Canning Ladle






--------------------
SpaceBaby
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Edited by SpaceBaby (05/30/21 04:06 PM)


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: SpaceBaby]
    #27329389 - 05/30/21 05:52 PM (4 months, 20 days ago)

Quote:

Josex said:
If you can find a clear enough bottle, sure, but good luck with that. Also I wouldn't trust plastic in a million years to grow an LC in.




:imwithstupid:


You can make anything work though, even if it's less than ideal.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


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OfflineSpaceBabyS
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27329629 - 05/30/21 10:37 PM (4 months, 19 days ago)

Quote:

Josex said:
Quote:

Josex said:
If you can find a clear enough bottle, sure, but good luck with that. Also I wouldn't trust plastic in a million years to grow an LC in.




:imwithstupid:


You can make anything work though, even if it's less than ideal.




Cool. It'll be one of my next projects. I have unmodded lids on tall 1/2-pints now. Not an ideal pour, but I wouldn't know another way to nocc jars from those. I'll [finally] look at a few lid teks. Here we go...:kittytakedown:

Ummm, don't you suggest plastic shaker bottles for your LC pours in a Josex's Poke TEK to avoid the whole lid-mod thing?

Injecting the poke into the Nalgene is certainly a vector but, if the LC is clean, then logic would seem to indicate that reducing the nocc to a sterile squeeze botttle nozzle in the same config that RR [and I] use for IPA would keep the bottle hand farther from the jar. But there remains the actual application to be performed and since this is my first LC project there is the greater possibility of contam caused by myc lack of practice. :shrug:

I saw the C typo; decided I like it. :mushroom2:


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SpaceBaby
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Re: The Biopsy Method: A different way of doing LC. [Re: SpaceBaby]
    #27330474 - 05/31/21 05:20 PM (4 months, 19 days ago)

Yes, I did recommend (years ago, I do things different these days) pouring into a blender bottle in the OP, but we have to make a distinction between growing LC from the start in a questionable container and pouring an already colonized liquid culture into another vessel to be used immediately.

If by any chance any bacteria were to land on your colonized LC while you're pouring, it won't likely represent a serious competition to an already established colony that you've used immediately to inoculate stuff.
On the other hand, if any bacteria were to make its way into the broth when you noc it up (or after because of faulty lid/container), you can bet your LC is doomed.


If what you're after is growing your LC in a container that is also good for pouring I'd look into media bottles. I think you'll find interesting mushboy's thread on LC, he explains this stuff there. The ones you posted won't let you see well, among other issues I see, potentially.

About plastic and threaded lids, go back a page or two, we were discussing it. You'll find there the reasons why I don't like plastic lids for LC (or anything plastic), but by all means try it out and let us know, you can make anything work.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


Edited by Josex (05/31/21 05:31 PM)


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OfflineSpaceBabyS
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27330764 - 05/31/21 10:48 PM (4 months, 18 days ago)

Quote:

Josex said:
Yes, I did recommend (years ago, I do things different these days) pouring into a blender bottle in the OP, but we have to make a distinction between growing LC from the start in a questionable container and pouring an already colonized liquid culture into another vessel to be used immediately.

If by any chance any bacteria were to land on your colonized LC while you're pouring, it won't likely represent a serious competition to an already established colony that you've used immediately to inoculate stuff.
On the other hand, if any bacteria were to make its way into the broth when you noc it up (or after because of faulty lid/container), you can bet your LC is doomed.


If what you're after is growing your LC in a container that is also good for pouring I'd look into media bottles. I think you'll find interesting mushboy's thread on LC, he explains this stuff there. The ones you posted won't let you see well, among other issues I see, potentially.

About plastic and threaded lids, go back a page or two, we were discussing it. You'll find there the reasons why I don't like plastic lids for LC (or anything plastic), but by all means try it out and let us know, you can make anything work.




Actually, that's a great answer for me. I like to understand stuff. I have 6 media bottle I use for agar. AND I think I forgot to put the agar into the agar for a couple. LOL :blush: But it seems that may work out well, hunh. I have a midi-fridge that's pure myco. I poured that one with the black on the rim - if the plates contam, it's ok. Anyway, of the remaining 5 media bottles in there, I think 3 may just be sterilized broth; if that's true I suppose I could PC the squirt bottles and one of the funnels and transfer in the SAB. But duh that adds a vector and I could just use the media bottles. :facepalm3: So never mind.

I have another clone, a GT. I am trying most to get canopies of the two clones and if other jars are successful, yay. ButI I want the clone flushes to test the strength of the clones. Now that I'm learning all these "mini-grow" TEKs I can test more clones without big harvests.

Actually, YOU probably could make anything work. I can gain the experience to eventually make anything work, probably.

Anyway, hopefully next time I remove the cap from the needle I won't biopsy myself. Shoulda noccd...nah.

:bow2:


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SpaceBaby
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27331873 - 06/01/21 10:30 PM (4 months, 17 days ago)

Forgive me if this has already been addressed. I've read in previous posts that some people have concerns about inoculating LC through directly through an injection port after doing a biopsy poke. In my opinion the following method allows this to be done safely and further reduces the risk of contamination by allowing you to keep the lid on the jar.

For this method you'll need an 18g 1 or 1.5" needle and a 22g 3.5" needle.



Insert the 18g needle into the injection port. This becomes your introducer needle.



Remove the stylet from the 22g needle and do your biopsy as usual.



Place the 22g needle through the 18g needle into the LC.



Replace the stylet to scrape out any mycelium in the lumen of the needle and remove the needle from the LC.



Remove the 18g needle from the injection port and you're done. It's a pretty slick technique which I've been using with great success.



Edited by Intrepid (06/01/21 10:31 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Intrepid]
    #27332879 - 06/02/21 06:31 PM (4 months, 16 days ago)

Cool! Interesting to see what people come up with.

I don't inoculate through SHIP's myself but iirc some people in this thread have had consistent success inoculating normally through a SHIP, from jars with foiled lids that are taken out from the PC directly to the SAB/flowhood.

It all comes down to doing what you're comfortable with (as long as it works) without breaking your piece of mind too much.

That large needle with that stylet thing?  First time I see that. What's that stylet  exactly? Is it like a blind needle?
And where can I buy something like that? Now that I see it I think I could use something like that for a particular thang.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex] * 1
    #27332920 - 06/02/21 07:11 PM (4 months, 16 days ago)

These are medical grade needles. The stylet fills the lumen so you don’t take out a plug of tissue while advancing it. I just withdraw the stylet slightly when using your biopsy technique. Lots of places sell them but here’s an example:

https://www.medofficedirect.com/b-braun-spinocan-spinal-needle-with-quincke-bevel-spinal-needle-22g-x-3-clear-hub-50-cs-12628.html?utm_source=google_shopping&utm_medium=ppc&gclid=EAIaIQobChMIkrrskYf68AIV5wutBh0pOAd0EAQYCCABEgLM6_D_BwE


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Re: The Biopsy Method: A different way of doing LC. [Re: Intrepid]
    #27332980 - 06/02/21 08:03 PM (4 months, 16 days ago)

Never seen that either, pretty cool.

I’ve been using silicone/rtv squirted over the hole as a ship for a while with no problems yet.


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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27347295 - 06/13/21 11:37 PM (4 months, 5 days ago)

Bump, great thread here! =)


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Re: The Biopsy Method: A different way of doing LC. [Re: Intrepid]
    #27347408 - 06/14/21 02:15 AM (4 months, 5 days ago)

Even without a stylet, the technique of using a big needle and a long thin needle is a good idea. When I inject a biopsy through a SHIP, it sometimes doesn't take. I can only assume the tissue stuck to the silicone. Using one needle inside another would solve this problem.


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