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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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Re: Pastywhyte's EzLC tek. [Re: bulbstoo]
#25885802 - 03/20/19 01:35 PM (5 years, 10 days ago) |
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I don’t add water to the agar plate simply because it’s not a large enough volume. I have done that in the past however and it does work. Also I’m using fewer syringes these days and simply pouring the water in. I also now usually just pour the LC into the recipient jars. Having a flowhood makes this a pretty easy process though I have done it in a sab as well.
I don’t use a stir bar for two reasons, first is that it would just get stuck in the cooled agar puck at the bottom and be limited in its usefulness. Second is simply that I have never used one and see no reason to start. Works fine without.
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Palmer1997
do€ob do£od
Registered: 03/16/19
Posts: 123
Last seen: 4 days, 11 hours
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#25948358 - 04/21/19 10:17 PM (4 years, 11 months ago) |
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Hey pasty, out of all the teks lc wise I’ve seen, this is the most simple and probably beginner friendly. It should come up when you click on LC in the main forums. Anyways, I don’t have much to spend on supplies but I could definitely get these jars and filters, and a spore syringe. Would it work if I inoculated it straight into the LC after sterilizing? I heard it’s not as great as using an agar wedge or what not.
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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Re: Pastywhyte's EzLC tek. [Re: Palmer1997]
#25948692 - 04/22/19 07:06 AM (4 years, 11 months ago) |
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While you probably could use a single drop of spore solution to start one of these, I would consider it a big risk. Agar wedge is the best way to start one of these.
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Palmer1997
do€ob do£od
Registered: 03/16/19
Posts: 123
Last seen: 4 days, 11 hours
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#25948833 - 04/22/19 09:47 AM (4 years, 11 months ago) |
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Thank you for replying, I’m so happy this site exists and veterans like you can help a noobstool like me, anyways I’m pretty hype, I’m going to be getting a Fiji strain cube, I was gunna get PE uncut but figured a more regular cube would be better for first time. I’m planning on doing agar work with malt dextrose, find some good monoculture (I think I have a good eye for it) then toss it in this upgraded LC and it should be pretty simple from there.
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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Re: Pastywhyte's EzLC tek. [Re: Palmer1997]
#25949196 - 04/22/19 02:02 PM (4 years, 11 months ago) |
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Don’t worry about getting isolates or monocultures. Just get a clean vigorous ms culture. You can always clone for consistently good genetics. With isolates it’s annoying because you need to fruit out at least 10 cultures to find anything good. Some isolates can’t even fruit.
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Palmer1997
do€ob do£od
Registered: 03/16/19
Posts: 123
Last seen: 4 days, 11 hours
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#25949284 - 04/22/19 03:08 PM (4 years, 11 months ago) |
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Oh wow I had no idea you always had to fully fruit each culture, I was under the impression you isolate until it’s all one uniform spread on agar, then fruit and work with that. But it makes sense that you must fruit out each transfer, I think that’s what you mean right? Or do you fruit out each transfer even if it has like 3 different sectors/strains?
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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Re: Pastywhyte's EzLC tek. [Re: Palmer1997]
#25949309 - 04/22/19 03:23 PM (4 years, 11 months ago) |
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No you don’t have to fruit every transfer. Don’t bother fruiting things that look weak.
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A2K
Traveller
Registered: 03/06/20
Posts: 6
Last seen: 2 years, 4 months
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#26668821 - 05/13/20 03:47 PM (3 years, 10 months ago) |
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Thanks much. Good info per usual.
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ALX8721
Registered: 03/06/19
Posts: 38
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#26808031 - 07/06/20 10:22 PM (3 years, 8 months ago) |
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Nice, reminds me of a “slurry tek” I found somewhere.
Basically using agar to ensure clean growth and then transfer the mixture of sterilized water and established culture into an LC jar, am I right? Def gonna try this out, thanks señor Pasty. Appreciate the write up!
-------------------- Just here to spread that love.
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sacredgeomancer
Ehh?
Registered: 01/20/18
Posts: 52
Last seen: 1 month, 12 days
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#27094156 - 12/17/20 01:16 PM (3 years, 3 months ago) |
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Pasty, thanks so much for the wonderful teks... You've helped me up my game significantly.
I just started two jars using the Super Ez LC method. I'm wondering if it would be feasible to transfer the final LC's to a Pyrex media bottle using a funnel (in a SAB).
Would this add too many new vectors? Would iso be enough to clean off the funnel before use, or would one want a pp5 funnel that can be PC'd? Does the pour method require you use all of your LC from an individual jar in one go?
Thanks much
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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If I was going to want the final colony in that Pyrex bottle it would be better to make it there in the first place. Not sure if you would want to do that but you could make the super EzLC in the bottle. But barring that I find that while it’s likely plausible that you could successfully transfer it, I consider it a fair bit of exposure.
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sacredgeomancer
Ehh?
Registered: 01/20/18
Posts: 52
Last seen: 1 month, 12 days
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#27094279 - 12/17/20 02:17 PM (3 years, 3 months ago) |
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Right on, thank you.
When you pour your LC to grain, are you using that jar to completion? I think it was mentioned in this thread that the lip of the jar becomes a vector after the pour. Just curious as to how you rock it.
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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With the super ez it’s like a G2G, you pour the whole thing all in the same session. That eliminates the risk.
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sacredgeomancer
Ehh?
Registered: 01/20/18
Posts: 52
Last seen: 1 month, 12 days
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#27094753 - 12/17/20 06:37 PM (3 years, 3 months ago) |
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the man
still masked
Registered: 08/12/99
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would pectin perhaps be better in this situation? perhaps little pectinase to loosin things up if wanna get crazy lol.
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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Re: Pastywhyte's EzLC tek. [Re: the man]
#27095020 - 12/17/20 09:51 PM (3 years, 3 months ago) |
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Quote:
the man said: would pectin perhaps be better in this situation? perhaps little pectinase to loosin things up if wanna get crazy lol.
Why would “loosening things up” be better?
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Tattersail
Registered: 04/11/18
Posts: 285
Last seen: 2 years, 1 month
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#27095219 - 12/18/20 02:52 AM (3 years, 3 months ago) |
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This looks like a great tek! I'm not keen on messing around with silicone and stuff for lids (I use polyfil for grain jar lids). Would it be ridiculous to use a circle of wool felt or acrylic felt in the lids and stick a syringe through that? If not then I'm happy with blenderless LI. I probably don't do enough jars to justify doing LC anyway, but I like the idea of fast colonisation.
-------------------- LAGM2021 Trades We may lose or we may win, but we'll never be here again
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Shakedown Street
Friend
Registered: 09/05/20
Posts: 383
Last seen: 22 days, 5 hours
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Re: Pastywhyte's EzLC tek. [Re: Tattersail]
#27124233 - 01/03/21 03:57 PM (3 years, 2 months ago) |
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I am thinking of trying your Super EZ LC tek, Pastywhyte.
Of 5 LC jars I have, 3 have mycelium that looks healthy. I just put samples from those 3 to agar. If they still look good after they grow out on the agar, I will want to inoculate grain jars and then regrow more LC.
But if you have to wait 7-10 days, I'm not sure how much this "speeds up" grain inoculation.
Right now I only have half-pint jars, so I will probably try to get taller jars before I do this since you said taller is better.
Edited by Shakedown Street (01/03/21 03:59 PM)
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,830
Loc: Canada
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When you get recovery on the grain in 12 hours as opposed to 2 days it’s faster. I’m not looking at the time it takes for me to get a culture off a plate into the grain, that could take months for all I care. What I’m concerned with is how fast the grain itself colonizes once the culture hits it.
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Shakedown Street
Friend
Registered: 09/05/20
Posts: 383
Last seen: 22 days, 5 hours
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
#27124270 - 01/03/21 04:19 PM (3 years, 2 months ago) |
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I see.
I also read about L.I., but I don't fully understand it yet. Which tek do you think would be more foolproof for a newbie, L.I., or your Super EZ LC tek ?
BTW, thank you for writing this tek.
Edited by Shakedown Street (01/03/21 04:20 PM)
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