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InvisibleMateja
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Re: Some notes about LCs [Re: Nobler Hino]
    #27053376 - 11/23/20 05:16 AM (3 years, 2 months ago)

Depends on what you mean by "pretty rich", iirc bod has measured the nutrient density in undiluted GW and it is actually weaker than the equivalent of 2% malt extract. This is further confirmed IME after noticing that my agar cultures are always stronger looking, faster growing on any kind of 2% recipe than on undiluted GWA. How myc looks on GWA in my experience reminds me of myc on 1% MEA.


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InvisibleNobler Hino
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Re: Some notes about LCs [Re: Mateja]
    #27053395 - 11/23/20 05:34 AM (3 years, 2 months ago)

That's interesting :takingnotes:


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"The sacred mushroom takes me by the hand and brings me to the world where everything is known. It is they, the sacred mushrooms, that speak in a way I can understand. I ask them and they answer me.”
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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Nobler Hino]
    #27053814 - 11/23/20 11:48 AM (3 years, 2 months ago)

It’s ghetto and weird but I still like my EzLCs. These are some grain water and instant coffee LC’s that are looking good to me. Gonna get em to grain soon.



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OfflineGeinstein
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Re: Some notes about LCs [Re: Pastywhyte]
    #27053828 - 11/23/20 12:01 PM (3 years, 2 months ago)

Quote:

Pastywhyte said:
It’s ghetto and weird but I still like my EzLCs. These are some grain water and instant coffee LC’s that are looking good to me. Gonna get em to grain soon.






Instant coffee?
I have actually been wandering about adding it now for a long time, just never had the guts to five it a shot.

At what % or what ever do you use it?


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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Geinstein]
    #27053874 - 11/23/20 12:38 PM (3 years, 2 months ago)

I can’t really say for this batch, it’s just what was added to the grain soak water. However I’ve used instant coffee in potato flake agar at 0.25% and the cultures seemed to do good on it.


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Invisiblemushhead
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Re: Some notes about LCs [Re: Pastywhyte]
    #27053894 - 11/23/20 12:58 PM (3 years, 2 months ago)

I've been working on my LC nute levels. Trying to get them as clear as everyone's here.
My LCs are clean, but have sediment because I've been experimenting to nute levels.
My first batch is 5g LME to 500ml distilled water. LOTS of sediment. 
Second is 2.5g LME to 500ml distilled water. Medium-minor sediment.
Third is 1g LME to 500ml distilled water. Mostly none. Small amount visible on bottom of jar.
Fourth is .5g LME to 500ml distilled water. Pretty clear.
Pics after I finish a bunch of errands today.


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Offlinestarbones
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Re: Some notes about LCs [Re: mushhead]
    #27053899 - 11/23/20 01:05 PM (3 years, 2 months ago)

Asked a question in another thread but this thread is more apt for it.

Let's say I have a box full of used 50ml BD syringes I've used for LCs before. I had a hairbrained idea about it..

Could I add ELME to boiling water in the right ratio then suck it up, squirt it out and repeat as if I was sterilizing the syringe and then on the final draw take up 50ml of the boiling water/ELME (The BD syringes can take another 10ml) and will that be sterile enough for LC? I've got to think that a 20 minute boil will kill any mold and bacteria present even with ELME in there.

Then when the syringe cools, could I swap a fresh needle on, suck back 10ml out of an LC master and then let it grow in the syringe with the new fresh nutrients?

What about two pots on the go, one with boiling water, one with boiling water + 2% ELME, suck and spit suck and spit from the plain boiling water, then on the final succ take up 40-50ml of boiling ELME? I can't see bacteria and molds surviving in it.

I'm gonna get fucked by endospores aren't I?
Edit: Wait fuck, if we're told we can sterilize with boiling water isn't that wrong since endospores will survive the trip?


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Edited by starbones (11/23/20 01:09 PM)


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Invisiblemushhead
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Re: Some notes about LCs [Re: starbones]
    #27053905 - 11/23/20 01:11 PM (3 years, 2 months ago)

Well you would want to sanitize your syringe with a couple of pulls of boiling water to clean it out proper.
You would have to squirt it out into the sink, not into your boiling water because it could contaminate the whole pot. There are heat-resistive endospores to worry about.
You would need to sterilize in the PC your nutrient liquid for the syringe as well beforehand.
After its been sterilized in your PC and your syringe as sanitized as it can get you could suck up a little LC and then some of your nute water and it could work.
But why go through all that when you could just expand an already clean LC?
Make multiple syringes you know are clean right off the bat like?
Correct me if I'm wrong though. But I read up on this a few months back.


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Offlinestarbones
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Re: Some notes about LCs [Re: mushhead]
    #27053917 - 11/23/20 01:15 PM (3 years, 2 months ago)

Yeah, I'm overthinking this a lot.
My bad, I wish I could just sterilize LC in a syringe in my PC without the risk of it boiling out, just so I could pull a little LC from the master with each one and have a drawer full of 50ml's colonizing ready to go.


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Listen, I'm steel fisted with the iron lung
Heavy metal ballads out the guitar where lions run.




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Invisiblemushhead
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Re: Some notes about LCs [Re: starbones]
    #27053938 - 11/23/20 01:27 PM (3 years, 2 months ago)

You can sterilize a syringe in the PC.
I have read that some wrap it in tinfoil, place it within a jar with a lid, and PC it for 15-20min to sterilize.
I am unsure if this works because I have access to a huge amount of sterile luer lock syringes for basically free. 18g needles and all.


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Offlinestarbones
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Re: Some notes about LCs [Re: mushhead]
    #27053964 - 11/23/20 01:41 PM (3 years, 2 months ago)

Oh I meant sterilizing the syringe WITH LC solution inside of it in the PC.

I've got plenty of 50ml syringes and more than enough needles I never reuse anything usually but what I was trying to do was take a small amount from a master sucked up into a syringe full of sterilized LC so I can have a bunch of LC syringes ready to go when knocking jars up. Instead of constantly refilling syringes during innoculations. Just use, toss, use, toss.
Instead of fill, use, change needle (I use a fresh 18g everytime I go into my LC quarts, cheap enough to not do the flaming) then fill, use, change needle, fill, use, blah.

I guess I could sterilize a bunch, make a new slave quart and fill them all prior to inoculation day.


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Listen, I'm steel fisted with the iron lung
Heavy metal ballads out the guitar where lions run.




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Invisiblemushhead
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Re: Some notes about LCs [Re: starbones] * 1
    #27053969 - 11/23/20 01:43 PM (3 years, 2 months ago)

I do the same thing except I have a bunch of ready sterile syringes at hand.
I choose one master jar and make a bunch of syringes, expand into a new master after making syringes.
Inoculate as needed.
So if you sterilized all your syringes without anything in them and then used them with clean needles for your LC it would achieve the same thing.


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InvisibleLemgrub
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Re: Some notes about LCs [Re: mushhead] * 1
    #27053976 - 11/23/20 01:45 PM (3 years, 2 months ago)

I don't see why you can't sterilize the nutrient water in the syringe in the PC, needle up, you shouldnt lose more than a couple mL of solution.

I wonder what the success rate would be with josex's poke method, but just suck the biopsy into the syringe rather than squirt it out. Could be a good way to build up a library of a bunch of distinct cultures.


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InvisiblePastywhyteMDiscord
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Re: Some notes about LCs [Re: Lemgrub] * 1
    #27053984 - 11/23/20 01:47 PM (3 years, 2 months ago)

The boiling water method to sterilize syringes is less effective at higher elevations. I’ve had some spotty results with it and now wrap and PC syringes (if I’m not lazy and just use new ones).


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Invisiblemushhead
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Re: Some notes about LCs [Re: Lemgrub]
    #27053993 - 11/23/20 01:50 PM (3 years, 2 months ago)

I did mention that it could be done in the PC.
Wrapped in foil and in a jar, I haven't ever done it that way so I couldn't say if it works or not.
I am sure that with the needle up, wrapped in foil, that it wouldn't loose much.
But wouldn't diffusion happen at a certain point, contaminating the syringe?


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Offlinestarbones
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Re: Some notes about LCs [Re: Lemgrub]
    #27053995 - 11/23/20 01:52 PM (3 years, 2 months ago)

You mean have sterilized LC in the syringe, poke the plate then attempt to suck the agar back in the needle? Wonder if it would clog.

Yeah that feels like it would work if everything could be kept sterile. Schlurp the agar snip back into the LC, cap the needle and let it grow.
I guess I'll try LC in a syringe upside down with the needle up to vent tonight. Thinking it would be best to let the PC cool slowly overnight.

Yeah I usually use brand new since the equipment isn't expensive but I noticed them sitting there and had a brainfart. Will try PCing them tonight standing plunger side down in jars since with the plunger withdrawn I can't seal a jar, just foil wrap.

If the LC stays in the syringe I'm going to try that josex poke idea.

Wait diffusion?



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Listen, I'm steel fisted with the iron lung
Heavy metal ballads out the guitar where lions run.




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OfflineA.k.aM
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Re: Some notes about LCs [Re: starbones] * 1
    #27054006 - 11/23/20 01:59 PM (3 years, 2 months ago)

I’ve sterilized syringes full of water in the pc a bunch of times, I don’t see why it wouldn’t work with lme added.

You could just squirt a couple drops onto the myc, scratch it up and suck it back into the syringe and it seems like it would grow in the barrel alright.


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Invisiblemushhead
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Re: Some notes about LCs [Re: A.k.a]
    #27054010 - 11/23/20 02:02 PM (3 years, 2 months ago)

Quote:

A.k.a said:
I’ve sterilized syringes full of water in the pc a bunch of times, I don’t see why it wouldn’t work with lme added.

You could just squirt a couple drops onto the myc, scratch it up and suck it back into the syringe and it seems like it would grow in the barrel alright.



Now I'm gonna have to try this.
diffusion is the process of CO2/O2 exchange through the plastics of the syringe. I have read that it could contaminate syringes over a long period of time.


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Offlinestarbones
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Re: Some notes about LCs [Re: mushhead]
    #27054055 - 11/23/20 02:20 PM (3 years, 2 months ago)

Well alright then it's settled, making sterilized LC solution syringes to quickly(?) expand healthy looking myc colonies from agar.

Same as prom night, open it up, poke, suck and let it spread inside the barrel.
Be really handy to have for the muda-esque bottles I've got going on and looking for good clones. Smart fuckers around here man, smarties.


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Listen, I'm steel fisted with the iron lung
Heavy metal ballads out the guitar where lions run.




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Offlinesonoramo
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Re: Some notes about LCs [Re: starbones]
    #27054105 - 11/23/20 02:51 PM (3 years, 2 months ago)

Quote:

starbones said:
Oh I meant sterilizing the syringe WITH LC solution inside of it in the PC.
...
Instead of fill, use, change needle (I use a fresh 18g everytime I go into my LC quarts, cheap enough to not do the flaming) then fill, use, change needle, fill, use, blah.

I guess I could sterilize a bunch, make a new slave quart and fill them all prior to inoculation day.




Are you thinking to grow mycelium inside the syringe? I'd worry about clogging the needle. Even after spinning a bar inside an LC, mycelium clumps often block the tip of my 16ga needles as I'm drawing into the syringe. Not a huge problem because I can push it back out and try again. If that happens with a clump that's growing inside the syringe, there won't be any backing out.


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