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 Ababyphoenix
You
 
Registered: 03/21/19
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 Re: Josex' Poke: A different way of doing LC.  [Re: Josex]
#27255324 - 03/16/21 08:39 AM (3 years, 1 month ago) |
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My first poke lc’s are showing some nice growth i think? What do u guys think? I havent an eye for lc yet but the waters clear not cloudy!
  
All the brown specs are lme that didnt dissolve
I didn't clean the jars very well before i used them for LC. some of them have my own hair floating in them cause i guess I'm nasty as fuck lol. Im guessing that wont be a problem since ofc its sterile. Right? 
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     Pho's MushiAventures:
 My '21 GLOG  LAGM 2022
Edited by Ababyphoenix (03/16/21 09:16 AM)
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 Josex
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Re: Josex' Poke: A different way of doing LC. [Re: Ababyphoenix]
#27255733 - 03/16/21 02:45 PM (3 years, 1 month ago) |
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They're probably fine but most times it's impossible to tell from a pic. Are you using LME, looks kinda cloudy for LME.
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Ababyphoenix
You
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Posts: 1,015
Loc: Human Body
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27255850 - 03/16/21 04:15 PM (3 years, 1 month ago) |
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Yeah lme but the lme didn’t dissolve properly and made it a but cloudy when shaken
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Pho's MushiAventures:
My '21 GLOG LAGM 2022
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: Ababyphoenix]
#27256747 - 03/17/21 06:09 AM (3 years, 30 days ago) |
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You've probably heard differing opinions on how to get a crystal clear broth but I'm going to tell you how I get there.
I use anywhere from 0.2 to 0.3% LME to water ratio, put the mix in a measuring jar, stir it to try to dissolve the LME as much as you can while it's cold, microwave the jar until it's hot, take it out of the MW and stir it some more, put it back in the MW and blast it until the first bubbles from boiling pop up.
Give it a try to see how it works for you.
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A.k.a
Stranger
Registered: 10/27/19
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27256882 - 03/17/21 08:20 AM (3 years, 30 days ago) |
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I recently made like 15 lcs with varying amounts of lme, but unfortunately didn’t measure it.
Most of the time I had just put a sprinkle of lme in and the broth would come out clear and colonize fine but never got super thick like pics I’d see. So I made some with way more lme and they came out dark yellow. It took like 3-4 weeks but most of them colonized nice and thick and eventually turned clear or close to it. It was kinda sketchy for a while cuz there was sediment and it’s cloudy so it’s harder to tell if it’s clean but in the end it made a much better lc.
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 LAGM2020      
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Ababyphoenix
You
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
#27257320 - 03/17/21 11:54 AM (3 years, 30 days ago) |
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Thank you both for the insight i appreciate it! Yeah i made a round of LC's using your tek Josex. This is my second attempt and i still cant get the LME to dissolve correctly. I must not have gotten the water hot enough but i used one of the electric coffee stirrers i use for agar as well and stirred the fuck out of it and they still came out cloudy and weird. Third times the charm, but i have a feeling these LC's are going to be fine just lots of undissolved LME.
The last batch of lcs i made the lids weren't good and i had an off feeling about them so i didn't inoculate and surely a week or 2 later and the water was cloudy and smelled rancid upon opening so. Im sure ill figure it out somehow. How do u guys test these LC's without innoc ports if you only open it once then pour it when the session is over. Id technically have to open the LC pour a bit onto an petri then re-cap which is a vector in itself. The once perfect lc could now test good but be bad. Maybe the move is to pour it to a plate and your masters in the same session then if the plate contams throw the jars? but then why even test it your already shooting yourself in the foot by not testing it really. Then there's the vector of silicone ports but making a syringe seems like... Idk i want as little vectors. Sorry yall I'm baked as fuck just got home from work. Il stop typing now 
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Pho's MushiAventures:
My '21 GLOG LAGM 2022
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tiptrippy
The Mechanic
Registered: 09/09/20
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Re: Josex' Poke: A different way of doing LC. [Re: Ababyphoenix]
#27265245 - 03/22/21 11:09 PM (3 years, 25 days ago) |
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I have 4 jars of LC here. Made with 1gLME/500ml distilled. I'm just looking for some experienced eyes here.
 
These first 2 pics are a Jambo clone. Noc'd on 3-16. It seems clear and healthy but looks more clumped together than the next few jars and idk if that is a concern or just a different species of myc.
This jar is APEU from spore print on agar also noc'd on 3-16
 
These 2 pics are AA+ from spore print on agar also noc'd on 3-16. This one looks to me like it is almost fully colonized and ready to pour but I haven't made it this far with LC yet so I'm a bit unsure.
And finally this last jar is an x7x clone I took from an agar dish pin. From the reading I've done about LC teks this one looks very cloudy which means bacterial and I should dump it. Yes?
Thanks for any advice.
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: tiptrippy]
#27265386 - 03/23/21 02:32 AM (3 years, 25 days ago) |
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I can agree with you on everything you said. The first one needs some swirling, pans don't clump for me in LC if I swirl regularly. The APEU one looks good and almost done. AA+ is ready. The last one is obviously taminated 
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tiptrippy
The Mechanic
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27265613 - 03/23/21 07:51 AM (3 years, 24 days ago) |
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That's great to hear I'm getting an eye for LC! Thanks for the confirmation I appreciate it
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Bsdgaou
Registered: 02/20/19
Posts: 340
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27280934 - 04/24/21 11:34 PM (2 years, 11 months ago) |
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Quote:
Josex said:
I can agree with you on everything you said. The first one needs some swirling, pans don't clump for me in LC if I swirl regularly. The APEU one looks good and almost done. AA+ is ready. The last one is obviously taminated <img src="https://files.shroomery.org/smileys/thumbdown.gif" alt="" title=""/>
Hey Josex. I found your tek here but I have some questions.
You said that use only once that LC... what does that mean? Just one poke from syringe and then no more? So if you have 20ml syringe, so only 20ml you take from LC?
Also don't fully understand where you suck up into syringe? As I understand, you don't like silicone ports. You open lids in front of flowhood or what?
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: Bsdgaou]
#27281068 - 04/25/21 04:15 AM (2 years, 11 months ago) |
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I said a lot of things in the original post that I don't do anymore (it is almost 4 years old).
I make LC's ranging from 150ml to 200ml and I don't pour anymore. I aspirate into 60ml syringes. I only aspirate what I'd need and leave the remaining LC in its jar to be used later on if I want. I use a modified lid for aspirating, no silicone port or self healing injection port.
Edited by Josex (04/25/21 04:50 AM)
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senseit
Listening
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27281458 - 04/25/21 11:42 AM (2 years, 11 months ago) |
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Quote:
Josex said:
I said a lot of things in the original post that I don't do anymore (it is almost 4 years old).
I make LC's ranging from 150ml to 200ml and I don't pour anymore. I aspirate into 60ml syringes. I only aspirate what I'd need and leave the remaining LC in its jar to be used later on if I want. I use a modified lid for aspirating, no silicone port or self healing injection port.
You mean just a lid with a filter on top right? What gauge needle do you use to aspirate?
Edited by senseit (04/25/21 11:43 AM)
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: senseit]
#27281463 - 04/25/21 11:46 AM (2 years, 11 months ago) |
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No, I use these lids.
  
I aspirate without needles and attach them right after aspirating. Use 14G needles.
Someday I'll make a tek on these lids (been saying that for ages now), they sure are handy.
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bodhisatta 
Smurf real estate agent
Registered: 04/30/13
Posts: 61,891
Loc: Milky way
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27281464 - 04/25/21 11:47 AM (2 years, 11 months ago) |
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Quote:
Josex said:
I said a lot of things in the original post that I don't do anymore (it is almost 4 years old).
I make LC's ranging from 150ml to 200ml and I don't pour anymore. I aspirate into 60ml syringes. I only aspirate what I'd need and leave the remaining LC in its jar to be used later on if I want. I use a modified lid for aspirating, no silicone port or self healing injection port.
Woah it really has been that long
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fahtster
Now With 33%More Faht
Registered: 06/17/06
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27281467 - 04/25/21 11:49 AM (2 years, 11 months ago) |
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Yeah man com’on! Is there a hole in the bottom (top) of the small glass jar for GE? Like, did you use a diamond bit to drill a hole in the glass? Do you have to use multiple syringes to get the air out from the tube? Looks gadgety.. love it lol
Faht
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senseit
Listening
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27281477 - 04/25/21 12:02 PM (2 years, 11 months ago) |
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Quote:
Josex said:
No, I use these lids.
I aspirate without needles and attach them right after aspirating. Use 14G needles.
Whoa those are wild!!! 
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: fahtster]
#27281505 - 04/25/21 12:29 PM (2 years, 11 months ago) |
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Quote:
bodhisatta said:
Woah it really has been that long
Yea time flies it's almost scary.
Quote:
fahtster said:
Yeah man com’on! Is there a hole in the bottom (top) of the small glass jar for GE? Like, did you use a diamond bit to drill a hole in the glass? Do you have to use multiple syringes to get the air out from the tube? Looks gadgety.. love it lol
Faht
Shrewd dude this Faht! 😂
Yeah, the jelly jar has a filtered hole, used a diamond bit.
You're always going to aspirate a tiny bit of air from the tube into the syringe, has never caused problems though. When you aspirate, that air is replenished with filtered air.
Sometimes I keep filled syringes around for days and they have always shown to be safe to use. Fuck I actually sell 10ml LC syringes (sometimes send them abroad) and nodody has complained yet.
Fairly recently I discovered that there's a commercial LC extractor lid (painfully expensive) that operates pretty much like my design, only that I think my design is superior 
I take the jelly jar out to inoculate through that poly filter with a 21G needle and squirt the biopsy into the broth. So these lids basically allow you to eliminate technique as plausible vector for contamination, not saying it can't happen, it's just pretty damn hard.
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fahtster
Now With 33%More Faht
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Posts: 9,378
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
#27281520 - 04/25/21 12:45 PM (2 years, 11 months ago) |
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Oh man.. promise I didn’t mean to come off shrewd.. prob shoulda put a “lol” after the com’on.. I just love your threads and write ups. Nothing but respect for ya.. I’m sure your syringes are great.. I was just trying to figure out your lid just from the pics and see if I got it right. I love it! 
Faht
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Josex
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Re: Josex' Poke: A different way of doing LC. [Re: fahtster]
#27281523 - 04/25/21 12:49 PM (2 years, 11 months ago) |
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I meant shrewd as clever man was j/k, I know you have good intentions haha
Thanks
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fahtster
Now With 33%More Faht
Registered: 06/17/06
Posts: 9,378
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Re: Josex' Poke: A different way of doing LC. [Re: Josex] 1
#27281535 - 04/25/21 01:05 PM (2 years, 11 months ago) |
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Ah. I had to look up shrewd... I thought meant something completely different haha maybe it was the “rude” sound in it
Faht
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