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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: micelio]
    #27215154 - 02/19/21 06:04 PM (7 months, 21 days ago)

It was probably the lack of space for shaking, didn't want to leave too much air in the syringes, anyway it's a crap method imo.
I had LC from 0.2% looking so thick in a proper vessel you couldn't even see the stir bar, so no, it couldn't be a lack of nutes because the broth in the syringes was also at .2% strength. I also did 2% lme in the past and tbh I couldn't see any advantage to doing that other than clouding the broth...not even faster leap off on grain that I could notice or faster colonization of the broth either, but to each their own.


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

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Invisiblemicelio
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27215329 - 02/19/21 07:51 PM (7 months, 21 days ago)

I certainly wouldn't argue with you...

I'm looking to do small batches with a fast turnaround.

That would be an interesting comparison test. Do a few bottles at .2% and a few bottles at 4%..


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OfflineA.k.a
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Re: Josex' Poke: A different way of doing LC. [Re: micelio]
    #27215336 - 02/19/21 07:55 PM (7 months, 21 days ago)

Lol you probably gotta suck the myc in and then suck some air straight from the hood in after.


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OfflineEdmunter
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
    #27228547 - 02/27/21 03:07 AM (7 months, 14 days ago)

All my test plates look ok i think.  I guess im looking at the outer edges for usual growth and foreign cultures.

What do you reccon.  The Ovoid culture really looked suspect as I think I added too much agar.  This is what bad cultures used to look like for me b4 adding agar.  Usually cultures collect together and kind of grow as one.



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OfflinetiptrippyS
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
    #27228609 - 02/27/21 05:11 AM (7 months, 13 days ago)

Quote:

tiptrippy said:
Quote:

On Fri Feb 26 2021 02:07 PM, tiptrippy said:
     




What do you all think about this jambo LC? Does it look healthy to you guys?

This is my first ever LC nocced on 2.14 with josex poke biopsy




Does this look healthy?


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OfflineA.k.a
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Re: Josex' Poke: A different way of doing LC. [Re: tiptrippy]
    #27228647 - 02/27/21 06:41 AM (7 months, 13 days ago)

Yeah that looks good to me. As long as the broth is clear you’re prob ok.


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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
    #27228657 - 02/27/21 07:04 AM (7 months, 13 days ago)

:whathesaid:

Lol Ed, a beer bottle that's pretty awesome. Must be good for pouring. Adding agar to the broth has a few important advantages imo, but on the other hand it clouds it a bit. I don't care though.


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

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Offlinemcnugget
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27228704 - 02/27/21 08:23 AM (7 months, 13 days ago)

These 2 Lc's were made 12 days ago.  One is cube and the other is lions mane.  I don't really see any growth in the cube jar, just some tiny particles which I'm thinking might be clumped LME.  The lions mane looks right to me as far as I can tell. 

These are my first 2 Lc's, both are made with 500ml distilled water and 2g LME.  Did the cube just fail to launch? 

This is the cube after the jar settled for about 30 mins.  The fine sediment in the jars settles after an hour or 2.



This is after a light swirl, you can see small particles on the left side.


This is the lions mane after sitting for 30 minutes.



And after a light swirl.



Any thoughts or input?  Going to fire up the pc today and give it another shot.


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OfflineEdmunter
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27230036 - 02/28/21 04:20 AM (7 months, 12 days ago)

I want to say yes! beer is the way I roll!!!! and it is, but these bottles are screw on lids and old apple cider vinegar bottles.  We get through 1 a week so I use them all..... I added a filter and a ship....


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OfflineEdmunter
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
    #27233228 - 03/02/21 09:10 AM (7 months, 10 days ago)

I had 2 lcs go bad one after another.  I think it might be when im shipping out to a syringe.


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Invisiblecoversall
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
    #27233249 - 03/02/21 09:35 AM (7 months, 10 days ago)

I had weird one recently. Super clear broth, nice myc growth in the LC, colonised like all my others. I think it was even made from the same plate as another LC that is knotting right now. I injected it into a pan-cake with all the ingredients I have used in the past, plus a small amount of ground coffee. The myc recovery was fast, and initial growth looked good, then boom. total stall out. The pots have sat for over a week with no growth at all.

Did the coffee kill it? I PC'd the pan-cakes for like >90 mins at 15 psi.

Did I just strike out and get a bum culture?

Reckon there was something lurking in the LC that wasn't visable?

I didn't test the LC on agar, because I'm lazy...

:kummeli:


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..:: E V E R Y  ::..

..:: New? Start here. ::..
..:: How I Panaeolus. From Agar to Tea ::..


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OfflineFrugalFungi
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Re: The Biopsy Method: A different way of doing LC. [Re: coversall]
    #27233484 - 03/02/21 12:50 PM (7 months, 10 days ago)

I just inoculated my first set of jars with my first lc attempt using the josex poke method! and the jump off took one day! Very exciting stuff. How does this test plate look? I think it looks good myself.


Edited by FrugalFungi (03/02/21 01:09 PM)


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InvisibleWall.E
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Re: Josex' Poke: A different way of doing LC. [Re: coversall]
    #27233882 - 03/02/21 05:48 PM (7 months, 10 days ago)

Quote:

coversall said:
I had weird one recently. Super clear broth, nice myc growth in the LC, colonised like all my others. I think it was even made from the same plate as another LC that is knotting right now. I injected it into a pan-cake with all the ingredients I have used in the past, plus a small amount of ground coffee. The myc recovery was fast, and initial growth looked good, then boom. total stall out. The pots have sat for over a week with no growth at all.

Did the coffee kill it? I PC'd the pan-cakes for like >90 mins at 15 psi.

Did I just strike out and get a bum culture?

Reckon there was something lurking in the LC that wasn't visable?

I didn't test the LC on agar, because I'm lazy...

:kummeli:




Doesn't bacteria cause pans to stall out sometimes? If you didn't test it then there's obviously no way of knowing whether it was the lc or not. Pan cultures are finicky too, but you know that, so idk


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OfflineEdmunter
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Re: The Biopsy Method: A different way of doing LC. [Re: FrugalFungi]
    #27234516 - 03/03/21 02:19 AM (7 months, 10 days ago)

Quote:

Hermansonking said:
I just inoculated my first set of jars with my first lc attempt using the josex poke method! and the jump off took one day! Very exciting stuff. How does this test plate look? I think it looks good myself.






Looks good atm.

Regarding testing im now filling a syringe and testing first.  I have more that 10 cultures on the go so patience is the key to success from now on


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Re: The Biopsy Method: A different way of doing LC. [Re: Edmunter]
    #27235128 - 03/03/21 01:49 PM (7 months, 9 days ago)

The lc smelled great when I was down aspirating I smelled the leftover. I'm really amazed at the different colonization speed with lc. It's been three days and I can see growth starting on 30% of my grains although it's weak and whispy looking.0


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OfflineBSUUF2
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Re: The Biopsy Method: A different way of doing LC. [Re: FrugalFungi]
    #27235175 - 03/03/21 02:27 PM (7 months, 9 days ago)

The mothership has landed...



2.5 liters water
25 g LME
2.5 g agar agar

Fresh out of the pressure cooker. Waiting to cool down and be inoculated... :mushroom2:


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OfflineAbabyphoenix
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27235427 - 03/03/21 05:46 PM (7 months, 9 days ago)

Did my first pokes to lc after waiting till i had a cutlure i wanted to go big w/.
Wish me luck yall, first LCs!


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:sporedrop:Pho's Mushi Aventures:sporedrop: : First Agar Journey:bow2:  Pho Conquors Agar

:buddha: LAGM2021 :buddha2:


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Invisiblecoversall
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Re: Josex' Poke: A different way of doing LC. [Re: Wall.E]
    #27235476 - 03/03/21 06:36 PM (7 months, 9 days ago)

Quote:

Myc_Hunt said:
Quote:

coversall said:
I had weird one recently. Super clear broth, nice myc growth in the LC, colonised like all my others. I think it was even made from the same plate as another LC that is knotting right now. I injected it into a pan-cake with all the ingredients I have used in the past, plus a small amount of ground coffee. The myc recovery was fast, and initial growth looked good, then boom. total stall out. The pots have sat for over a week with no growth at all.

Did the coffee kill it? I PC'd the pan-cakes for like >90 mins at 15 psi.

Did I just strike out and get a bum culture?

Reckon there was something lurking in the LC that wasn't visable?

I didn't test the LC on agar, because I'm lazy...

:kummeli:




Doesn't bacteria cause pans to stall out sometimes? If you didn't test it then there's obviously no way of knowing whether it was the lc or not. Pan cultures are finicky too, but you know that, so idk




yeah to be honest I think I've been getting lucky with my LCs and I was so unsure about the culture I chose to  make these out of. It could well be that this LC was a bit gross.
Luckliy the other LC seems to be viable so I should have a harvest to print from :smile:


--------------------

..:: E V E R Y  ::..

..:: New? Start here. ::..
..:: How I Panaeolus. From Agar to Tea ::..


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InvisibleJessicaRabbit88
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27236128 - 03/04/21 12:33 AM (7 months, 9 days ago)

I'm just not having good luck with the LC. I love the Josex Poke method and have successfully used it a year ago when I first started. I suspect the problem is in my cultures... my grains are getting the dreaded grey despite my LC looking clear (made 2/17 and 2/22):



First unshaken and shaken


Second unshaken and shaken


Third unshaken and shaken (questionable)


Fourth unshaken and shaken


Here are the last agar transfers. Anything worth transfering? Last petri is toast...




I just want to get rid of this contamination.

My plan is to make a few more transfers and take grain samples from the best perfoming colonized jars and put them to agar petris...


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Invisibleshadyshroomie
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Re: Josex' Poke: A different way of doing LC. [Re: JessicaRabbit88]
    #27251344 - 03/13/21 03:11 PM (6 months, 30 days ago)

Started 2/14:



Started 2/23:



Both clones.

When mixing, how bad is it for some broth to touch the lid? The lid has a SHIP and .22micron syringe filter.
I know the broth should never touch a WHATMAN filter but how about these?

The broths were made with I believe 1g LME to 500mL water, although the one started on 2/23 I believe was slightly less


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Storing Cultures

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