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 Josex
#cheat_code
Registered: 11/13/15
Posts: 8,996
Loc: 
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 Re: Josex' Poke: A different way of doing LC.  [Re: micelio]
#27215154 - 02/19/21 04:04 PM (3 years, 1 month ago) |
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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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micelio
Song of Silence
Registered: 04/22/14
Posts: 1,547
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27215329 - 02/19/21 05:51 PM (3 years, 1 month ago) |
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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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 A.k.a
Stranger
Registered: 10/27/19
Posts: 16,966
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Re: Josex' Poke: A different way of doing LC. [Re: micelio]
#27215336 - 02/19/21 05:55 PM (3 years, 1 month ago) |
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Lol you probably gotta suck the myc in and then suck some air straight from the hood in after.
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 LAGM2020      
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Edmunter
Mr
Registered: 05/01/13
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
#27228547 - 02/27/21 01:07 AM (3 years, 1 month ago) |
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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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tiptrippy
The Mechanic
Registered: 09/09/20
Posts: 1,131
Loc: United States 
Last seen: 1 year, 8 months
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
#27228609 - 02/27/21 03:11 AM (3 years, 1 month ago) |
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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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A.k.a
Stranger
Registered: 10/27/19
Posts: 16,966
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Re: Josex' Poke: A different way of doing LC. [Re: tiptrippy]
#27228647 - 02/27/21 04:41 AM (3 years, 1 month ago) |
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Yeah that looks good to me. As long as the broth is clear you’re prob ok.
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LAGM2020
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Josex
#cheat_code
Registered: 11/13/15
Posts: 8,996
Loc:
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
#27228657 - 02/27/21 05:04 AM (3 years, 1 month ago) |
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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.
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mcnugget
Stranger
Registered: 06/22/13
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27228704 - 02/27/21 06:23 AM (3 years, 1 month ago) |
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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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Edmunter
Mr
Registered: 05/01/13
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27230036 - 02/28/21 02:20 AM (3 years, 1 month ago) |
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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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Edmunter
Mr
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
#27233228 - 03/02/21 07:10 AM (3 years, 1 month ago) |
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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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coversall
إِنْ شَاءَ ٱللَهُ
Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: Josex' Poke: A different way of doing LC. [Re: Edmunter]
#27233249 - 03/02/21 07:35 AM (3 years, 1 month ago) |
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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...
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FrugalFungi
Loafter
Registered: 10/12/19
Posts: 130
Last seen: 2 years, 20 days
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Re: The Biopsy Method: A different way of doing LC. [Re: coversall]
#27233484 - 03/02/21 10:50 AM (3 years, 1 month ago) |
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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 11:09 AM)
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Wall.E
Bacteria's Bitch
Registered: 06/05/20
Posts: 2,860
Loc: Fungal Void
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Re: Josex' Poke: A different way of doing LC. [Re: coversall]
#27233882 - 03/02/21 03:48 PM (3 years, 1 month ago) |
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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...
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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Life’s shit, but I’m loving it
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Edmunter
Mr
Registered: 05/01/13
Posts: 5,699
Last seen: 30 days, 13 hours
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Re: The Biopsy Method: A different way of doing LC. [Re: FrugalFungi]
#27234516 - 03/03/21 12:19 AM (3 years, 1 month ago) |
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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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FrugalFungi
Loafter
Registered: 10/12/19
Posts: 130
Last seen: 2 years, 20 days
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Re: The Biopsy Method: A different way of doing LC. [Re: Edmunter]
#27235128 - 03/03/21 11:49 AM (3 years, 1 month ago) |
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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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BSUUF2
derails threads
Registered: 10/15/20
Posts: 666 
Loc: not that important
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Re: The Biopsy Method: A different way of doing LC. [Re: FrugalFungi]
#27235175 - 03/03/21 12:27 PM (3 years, 1 month ago) |
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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... 
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LAGM2022
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Ababyphoenix
You
Registered: 03/21/19
Posts: 1,015
Loc: Human Body
Last seen: 1 month, 25 days
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27235427 - 03/03/21 03:46 PM (3 years, 1 month ago) |
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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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     Pho's MushiAventures:
 My '21 GLOG  LAGM 2022
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coversall
إِنْ شَاءَ ٱللَهُ
Registered: 06/06/20
Posts: 2,749
Loc: संसार
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Re: Josex' Poke: A different way of doing LC. [Re: Wall.E]
#27235476 - 03/03/21 04:36 PM (3 years, 1 month ago) |
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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...
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 
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JessicaRabbit88
Loveable Nerd
Registered: 09/16/19
Posts: 78
Loc: Far Far Away
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27236128 - 03/03/21 10:33 PM (3 years, 1 month ago) |
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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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shadyshroomie
MycoCurious
Registered: 10/29/20
Posts: 132
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Re: Josex' Poke: A different way of doing LC. [Re: JessicaRabbit88]
#27251344 - 03/13/21 01:11 PM (3 years, 1 month ago) |
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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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