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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20762843 - 10/28/14 07:47 AM (9 years, 7 months ago) |
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Oh shit. Ha ha. I am tired. Vacation wore me out
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: azur]
#20762932 - 10/28/14 08:33 AM (9 years, 7 months ago) |
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What? Vacation? Dude, you were hanging with me every day for the past week. We're neighbors. Boy, are you feeling okay, azur?
Also since I can't really tell if my LC is contaminated and each time I uncover it to suck some out introduces another avenue for contams, what's the best line to take to use it? Suck up all of it into multiple syringes at once, test a little bit of them, and then if it's all good, use the rest?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,885
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20762991 - 10/28/14 08:58 AM (9 years, 7 months ago) |
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Use a big syringe. I like 60cc myself. Fill 4 of them up, test each and then good to go.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Pastywhyte]
#20763150 - 10/28/14 09:55 AM (9 years, 7 months ago) |
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Sounds good. I still need to buy syringes. What's the preferred method of testing for contams? Agar?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,885
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20763185 - 10/28/14 10:01 AM (9 years, 7 months ago) |
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Agar is a pretty fast way to check. Brf and grain jars are ok too but I like agar as its really easy to spot anything not right. If you go with agar do a few plates tho just to be sure.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Pastywhyte]
#20763200 - 10/28/14 10:05 AM (9 years, 7 months ago) |
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Yeah for sure. My sterile technique isn't excellent yet and I'd hate to have a false negative on 1 test. If I do 10 agar plates and there's an even split between contam/safe, that means the LC is safe for sure then right?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,885
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20763287 - 10/28/14 10:29 AM (9 years, 7 months ago) |
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If you do ten plates and all ten turn out good, I would say your good. Otherwise your just rolling dice.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Pastywhyte]
#20763307 - 10/28/14 10:34 AM (9 years, 7 months ago) |
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I'm confused. I don't understand how I can get any clean plates without the entire syringe being good. It's definitely possible to get bad plates from a good syringe due to improper handling and poor sterilization techniques, but how would I get clean agar cultures without a totally clean LC?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,885
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20763333 - 10/28/14 10:40 AM (9 years, 7 months ago) |
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Depends on a few things. Mold spores and bacterial endospores take time to germinate. Even after they do it will take them sone timw to take over. If you have a few endospores from poor sterilization or bad inoculation practices its possible thqt you could have a few come up clean. A week later tho while your test plates are growing out could mean the entire LC is comprised. There is no mostly clean with LC. Either it is and every test is successful, or it isn't.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Pastywhyte]
#20763343 - 10/28/14 10:44 AM (9 years, 7 months ago) |
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Hm, okay, I guess that's the risk with LC and why a lot of people skip it. I feel like it's good to know and practice with though, especially when I can noc up a bunch of agar plates, grain jars, whatever with it. Thanks for the help, Yung Paste.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,885
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20763355 - 10/28/14 10:48 AM (9 years, 7 months ago) |
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LC has lots of upside just be smart how you use it
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Pastywhyte]
#20763367 - 10/28/14 10:53 AM (9 years, 7 months ago) |
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I'm injecting it straight into my veins LET'S GET THIS PARTY STAAAARTED. Na, it's hanging out on my fridge right now next to my Pasty plates.
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Munchauzen



Registered: 06/22/11
Posts: 14,346
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20764480 - 10/28/14 03:55 PM (9 years, 7 months ago) |
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Quote:
stareatclouds said: Haha, no worries (you f'ing piece of shit), but I think you're also confusing me with someone else with regard to that stirrer thread. Go drink some coffee, azur.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: Munchauzen]
#20784640 - 11/02/14 03:49 PM (9 years, 7 months ago) |
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I somehow missed the part about storing this in complete darkness so it's been out in the open for like 5 days. The area it'll be in is probably like 65F at all times (kitchen pantry closet) but will be dark. It should be fine right, just delayed growing?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,893
Loc: Milky way
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20784743 - 11/02/14 04:17 PM (9 years, 7 months ago) |
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Light is good at all stages. If something says to keep it in the dark you know its dated old bs info
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Re: A *beginner friendly* method for making Liquid Culture [Re: bodhisatta]
#20784917 - 11/02/14 05:09 PM (9 years, 7 months ago) |
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That's what I figured, but wasn't sure. I'll just keep it wherever since I don't think it matters much and I won't need the LC for a while.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: A *beginner friendly* method for making Liquid Culture [Re: stareatclouds]
#20785014 - 11/02/14 05:40 PM (9 years, 7 months ago) |
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My enoki LC got direct sun light for most of the day while it was colonising and it did fine.
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_OttO_
Over Stimulated



Registered: 06/01/05
Posts: 2,594
Loc: Up Over
Last seen: 12 days, 19 hours
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Re: A *beginner friendly* method for making Liquid Culture [Re: cronicr]
#20790549 - 11/03/14 10:44 PM (9 years, 7 months ago) |
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Quote:
cronicr said: ok i got two lc's ready to test out, soaking grains in the morning an i will update monday or so
Any updates?
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 5 months
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Re: A *beginner friendly* method for making Liquid Culture [Re: _OttO_]
#20790797 - 11/04/14 12:11 AM (9 years, 7 months ago) |
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yeah sorry guys felt like shit for awhile but indeed both test jars were fine spawned both to coir and even made another glc from both got 20 cc's of each to use so i'm gonna knock some jars up asap
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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blackout


Registered: 07/16/00
Posts: 5,266
Last seen: 7 months, 19 days
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Re: A *beginner friendly* method for making Liquid Culture [Re: cronicr]
#20792436 - 11/04/14 01:34 PM (9 years, 7 months ago) |
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Quote:
stareatclouds said: I have an old glass jar with a thick plastic lid. Does the lid HAVE to be metal?
Plastic can go soft & floppy when hot, so be careful about having the lid on tight, or tightening it afterwards. It can warp out of shape and when it goes cold it can hold this deformed shape.
Quote:
PussyFart said: No print/syringe is ever 100% clean, as the cap was most likely fruited in open air.
Do any vendors claim to have guarantee clean syringes? or say they go above & beyond normal methods. The most trustworthy syringes I have made were from small jar grows, where the jar was never opened after sterlization. You let a shroom grow inside the jar and when its about to drop spores you leave the jar on its side and get spores on the side of the jar. You can then inject water into the jar and suck out spores. I expect some spore producers would have extra careful methods.
If you grow on "soft agar" LCs you can visually inspect it better, this is agar made with 1/10th the amount of gelling agar. I was trying this method to produce "mycelia mats" to be harvested for consumption, but noticed it was still liquid enough to be sucked up into a syringe. I also thought this may work well when injected on grains, coating them in an agar mix, but it may invite contams.
I also wondered about making a floating agar plate in an LC jar. So you have a floating bottle cap with solid agar floating in an LC, you inject into the jar into the floating dish, once you see that it looks OK you shake up the jar so the agar falls into the LC. So you are minimising the need to transfer it, and no need for glove boxes or similar. There are other similar methods to this, e.g. if you had a test tube siliconed to the the inside wall of a jar it could have the solid agar in it. Or a mini tray of rice or grains could be used if people have trouble finding agar, or a mini PF cake. I also wondered if you had a layer of verm on the top would it just float and be easier to inspect.
I still think aeration is very important, after boiling most gases are driven out of solution. For LCs I use jars like this, but with a much longer silicone tube with a filter on the end. This allows you to pinch shut the tube and shake the jar like crazy to aerate it and break up myc.

I also use 4mm ID silicone tube which is quite long and has a metal plug in the end. This allows me to effectively pour from one jar to another, and thick myc can easily transfer through the relatively large diameter tube. I could be adding up to 200ml of LC directly to a growing chamber. I colonise directly in the actual grow chamber, so the grains are shaken only 1 time ever, after the LC goes in. It can sometimes be cased in just a few days after the LC goes in. This also reduces the need for mulitple injections and the contam risks that go with them.
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