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Offlineshroomshine
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #26880589 - 08/14/20 09:02 PM (1 year, 3 months ago)

I love that shit! my favorite TEK on here! Thanks man :thumbup::thumbup::thumbup:


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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: shroomshine]
    #26880705 - 08/14/20 10:10 PM (1 year, 3 months ago)

Thanks man. It's def my favorite tek too!
:awethumb:


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Invisiblemicelio
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #26880730 - 08/14/20 10:27 PM (1 year, 3 months ago)

This Is my tool for doing the poke....

I use a flamed ram rod with my blunt luer lock.

https://www.shroomery.org/forums/showflat.php/Number/26835799#26835799


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Re: Josex' Poke: A different way of doing LC. [Re: micelio]
    #26880738 - 08/14/20 10:34 PM (1 year, 3 months ago)

That's kewl man. For the record, I never ever liked squirting water on agar, hate it actually. I've got my trusty escalpel for transfers. The times I did squirt water on agar was only to fuck around testing cultures.


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Invisiblepablokabute
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #26881574 - 08/15/20 02:20 PM (1 year, 3 months ago)

Now that Im getting a bit better at agar works, Im gonna give this one a go. Sounds like a freeeakin good way to expand colonies!

Thanks mang.


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OfflineGeinstein
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #26905833 - 08/29/20 01:02 PM (1 year, 3 months ago)

Its probably been asked but can you poke through your port?
Will tissue still be transferred?


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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: Geinstein]
    #26905849 - 08/29/20 01:11 PM (1 year, 3 months ago)

Yes it will, but I can't recommend something I wouldn't do myself. Poking an unsterile port is not good practice, but if you insist and do it regardless, just make sure to test the LC on agar first before using it. You might get away with it and if it fails it won't matter much, it's just a bit of liquid you noc'd via poke, easy to make gallons if you want.


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OfflineGeinstein
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #26906041 - 08/29/20 03:42 PM (1 year, 3 months ago)

Aaaaaah
The ports are sterile fresh out of the the PC and these ones can be flamed so should be safe?
Thanks
Was scared tissue will be lost on the port somehow


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Offlinekarri0n
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27003551 - 10/25/20 10:07 PM (1 year, 1 month ago)

I just used this method for the first time today.

I used water from my oat boil for the broth, pc'd for 90 minutes. This shit is brown and not clear.

The transfer was from a clean T1 plate on a really cool looking rhizo


I did the squirt in a SAB, and had the LC jar open for probably less than 1 second.

Come to think of it, the squirt might have hit the glass instead of the broth. I didn't notice for sure, so I hope not.

The broth is not even a little bit clear, so I am looking forward to the possibility of it clearing up as the myc colonizes it.

I am still working on my sterile technique, but the motions felt pretty smooth and "good".

This will be the first time making LC and I am looking forward to having something for fast colonization times.


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Invisiblecoversall
إِنْ شَاءَ ٱللَهُ


Registered: 06/06/20
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Re: Josex' Poke: A different way of doing LC. [Re: karri0n]
    #27028793 - 11/08/20 05:07 PM (1 year, 26 days ago)

Quote:

karri0n said:
Come to think of it, the squirt might have hit the glass instead of the broth. I didn't notice for sure, so I hope not.





Hah! Just packed away my SAB after my first go at this and have the same thoughts going round my head. Suddenly realised that I didn't make sure it went straight to the broth!

Oh well, time will tell. Next time round that will be a priority!

:babypalm:


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OfflineCelestialexplorer1
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Re: Josex' Poke: A different way of doing LC. [Re: coversall]
    #27028824 - 11/08/20 05:30 PM (1 year, 26 days ago)

Everytime I make LC I wipe the port down with iso inside the sab and draw up two mls of LC and then scrap up some myc with a sterile first time use needle and squirt it through the wipes down port. I never take the lid off at all until I’m done with the LC


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Edited by Celestialexplorer1 (11/08/20 05:35 PM)


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Offlinebongoman
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Re: Josex' Poke: A different way of doing LC. [Re: Celestialexplorer1]
    #27054712 - 11/24/20 12:34 AM (1 year, 11 days ago)

How many times do you reuse the same needle? I imagine that flaming the same needle would blunt and degrade the sharpness quite quickly.

A new needle for every few pokes?


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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: bongoman]
    #27054757 - 11/24/20 01:08 AM (1 year, 11 days ago)

I used to change the needles for every sesh every time, never reused them.
Got lazy over time and currently I find myself often using needles that have had quite a few uses. No issues, used needles do the job just fine unless they're in really bad shape.

Just don't go overboard when flaming and be careful not to burn the hub at the base of the needle, that should extend its life. As soon as the needle gets red hot is done, and that takes 1 second or less with a proper torch.

But anyways, needles are dirt cheap if you know where to buy them, so you can use a new one everytime if that gives you more peace of mind.


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Invisiblecoversall
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27054858 - 11/24/20 04:38 AM (1 year, 11 days ago)

Well the two LCs I nocd with the needle poke look great (at least to my inexperienced eyes)!

Great tek!


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OfflineAdrenognomeS
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Re: The Biopsy Method: A different way of doing LC. [Re: coversall]
    #27064303 - 11/30/20 12:24 PM (1 year, 5 days ago)

Can you do this with say 28 gauge needle? What’s the smallest gauge that will work? Anybody have experience?


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Re: The Biopsy Method: A different way of doing LC. [Re: Adrenognome]
    #27064323 - 11/30/20 12:34 PM (1 year, 5 days ago)

No idea, it may work. Please try it and report back.


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OfflineAdrenognomeS
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27064357 - 11/30/20 12:59 PM (1 year, 5 days ago)

I can’t help but think transferring to plates a few times like this might find an isolate


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Offlinebongoman
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Re: The Biopsy Method: A different way of doing LC. [Re: Adrenognome]
    #27064787 - 11/30/20 04:39 PM (1 year, 4 days ago)

What percentage of pokes don’t lead to any growth? I just had one that showed nothing after a week in the broth. Is this unusual?


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: bongoman]
    #27064810 - 11/30/20 04:46 PM (1 year, 4 days ago)

It happens yes. It's not usual at all but I'd say 1 out of 20.

Poke it more than once if you want, no problems with that.


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Offlinebongoman
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27064837 - 11/30/20 05:04 PM (1 year, 4 days ago)

Thanks Josex. This is such a great method.

I suspect this is probably answered elsewhere in the thread but how important is stirring/agitation with LME-based broth? I don’t have a stir-plate and can manually swirl a few times a day I guess.

Also, do you get a clean pour from a mason jar when pouring into grain jars? I’ve been using media bottles for LC which pour nicely but need to keep some aside for agar prep and so will try out mason jars as my next LC containers. I would have assumed that mason jars give a “sloppy” pour.

Also, for some reason I feel tempted to ignore the test step where you squirt to agar and then grow it out then poke and squirt to the broth - simply my impatience I guess. How important is this step in your view?


Edited by bongoman (11/30/20 05:08 PM)


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