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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: Edmunter]
    #23856938 - 11/22/16 10:55 AM (7 years, 2 months ago)

No less than 45 min at 15 psi. I might up that tho cause I had a run where some of my DFA plates went bacterial. Could have been a temp spike that helped it happen but I don't like that being a potential vector.


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OfflineEdmunter
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23856952 - 11/22/16 11:01 AM (7 years, 2 months ago)

Good call, But can I put it in with some rye jars without destroying the sugar content.  I was reading about this today and its new to me.


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: Edmunter]
    #23857100 - 11/22/16 11:53 AM (7 years, 2 months ago)

Depends on the type of dog food and if you use simple sugar in the recipe. My dog food is mostly protein and no grain so when I make it I use grain water or brf to pump it up without risking caramelizing the media.


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23857207 - 11/22/16 12:34 PM (7 years, 2 months ago)

I've been doing my cheap generic dog food for about 45 min at 17 psi with no pour and haven't seen any contam in my test plates.
I'v always done 45 min with all media in no pour but I may lower my times with pda and mea next time.


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Re: Pastywhyte's EzLC tek. [Re: tombosley8]
    #23877752 - 11/29/16 10:03 AM (7 years, 1 month ago)

So after my last attempt which finally turned into some ind of Welsh beer im back!!!!

This PE culture is looking good and a few more days should do it before adding water.

DFA ratio 10:10 pressure cooked for an hour this time.  I should have made it less firm but forgot.  Lets see what happens.



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OfflineLobi
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Re: Pastywhyte's EzLC tek. [Re: Edmunter]
    #23953808 - 12/24/16 10:00 AM (7 years, 1 month ago)

:rockon:
I forgot to picture my results from this method but I tried it out and it kicked it ass. Perfect for trying out a new isolate or strain on the quick while waiting for masters to colonize with agar.


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InvisibleTeemo 6T3
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23953828 - 12/24/16 10:11 AM (7 years, 1 month ago)

Can i use only this?



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Re: Pastywhyte's EzLC tek. [Re: Teemo 6T3]
    #23953836 - 12/24/16 10:13 AM (7 years, 1 month ago)

polyethersulfone is autoclavable. It should work :thumbup:

A lot of people use syringe filters for LCs.
BTW. It looks like .2 syringe filters do not provide enough GE for grain jars. I have seen reports of stalled jars with .22 syringe filter lids.


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Re: Pastywhyte's EzLC tek. [Re: enlightenment]
    #23953849 - 12/24/16 10:21 AM (7 years, 1 month ago)

Good to hear, thanks bud :dancer:


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23957601 - 12/26/16 02:03 AM (7 years, 1 month ago)

Possible improvement on your method :
you can lower the agar % even more but add a lot more mix in the jar,
mycélium will still form a mat on the surface for inspection, and it can be shaken nearly the same way as a liquid.
No need to inject water then, and you removed a contam vector.

Possible drawback: the exact agar % in the mix will be critical to get that result and everyone will need to adapt the recipe slightly.


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: Chk] * 2
    #23957835 - 12/26/16 08:31 AM (7 years, 1 month ago)

Sounds like a pain to me. Aspirating really thick LC is hard to do sometimes, what you propose would be far thicker.

I must ask if you have tried it yet? Hard to improve on something if you haven't tried it. Give one of the methods a shot and see what you think. Personally I don't see pouring the water to be a real vector. It's the same as doing G2G.


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte] * 1
    #23957838 - 12/26/16 08:32 AM (7 years, 1 month ago)

Quote:

Aspirating really thick LC is hard to do sometimes, what you propose would be far thicker.




:whathesaid:


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23958489 - 12/26/16 02:32 PM (7 years, 1 month ago)

I didn't try your method TBH, but i will do.
but i already tried LC with very low agar %, didn't have much problem to suck it with a 14 gauge.


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Re: Pastywhyte's EzLC tek. [Re: Chk]
    #23960497 - 12/27/16 12:58 PM (7 years, 1 month ago)

Quote:

Chk said:
but i already tried LC with very low agar %, didn't have much problem to suck it with a 14 gauge.



same here, I had no troubles. People having trouble with "thick" LCs mean thick with growth, large strands get caught in the needle and cannot be sucked up, they are physically clogged. If I get a clog I just shoot it back out again, I have also used sharpened needles.

Here are some posts, on refers to it as "soft agar" and the later one as an "agar broth". Some liked how it coated the grains.

https://www.shroomery.org/forums/showflat.php/Number/7680529

https://www.shroomery.org/forums/showflat.php/Number/13781184/fpart/all/vc/1


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: blackout]
    #23960606 - 12/27/16 01:54 PM (7 years, 1 month ago)

If the agar gells enough to get 2 dimensional growth it's going to be hard to aspirate. If it's not gelling enough to get 2 dimensional growth then it's just plain LC. If you want to make plain LC then do it. This tek is for people who are not looking to make traditional LC.


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23960661 - 12/27/16 02:21 PM (7 years, 1 month ago)

Quote:

If the agar gells enough to get 2 dimensional growth it's going to be hard to aspirate




Not sure i agree, actually it can be nearly liquid, only a slight change in viscosity will allow the formation of a mat, and it can be like a soft gel, and still be easy to suck.

As said blackout above, it's clogging that pose a problem (even with a rather big needle), same experience here.


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Edited by Chk (12/27/16 02:22 PM)


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: Chk]
    #23960700 - 12/27/16 02:43 PM (7 years, 1 month ago)

Sounds good. I will replace the OP with the links provided :thumbup:


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23961410 - 12/27/16 09:26 PM (7 years, 1 month ago)

Quote:

Pastywhyte said:
This tek has been removed due to critical flaws pointed out by more knowledgeable people who didn't even need to try it. Refer to the links in the quote below. Sorry to waste your time.





Are you for real?
I was finally prepared to follow this tek and now it's gone


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Re: Pastywhyte's EzLC tek. [Re: ComebackKid]
    #23961451 - 12/27/16 09:57 PM (7 years, 1 month ago)

I am very "for real". If I have written this so poorly that people think it's about adding 0.5% agar to an LC to make a culture form a mat on the surface and that will be enough to spot satellites or bacteria colonies, then I have failed. If a TC can't understand the point of being able to trap media in a puck to allow for sediments to be avoided no matter what media is used, then it needs to be re written, maybe with more pics. If people think it's cool to come into a thread discussing a new method with links to old methods then perhaps it's superfluous to begin with.


I may repost this later once it's more fleshed out and made idiot proof. In its current form it's obviously too complex. Maybe the food coloring is too distracting. Or I might just delete most of my other writeups and stop posting my methods.


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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #23961519 - 12/27/16 10:46 PM (7 years, 1 month ago)

You are one of the most prolific and helpful posters around here, Pasty. Chill, and please repost the original Tek, or perhaps you could PM me.... Don't let others misunderstanding the message silence the messenger. Be easy, hope you continue to share your knowledge and inspire the rest of us.


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