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Fungus-Amongus
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Registered: 08/26/11
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GLC Questions
#15665849 - 01/14/12 03:24 PM (12 years, 5 months ago) |
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So I have a few questions regarding making a GLC. I would be following Agar's tek since it seems to be the only one I can find. I understand that using a grain jar is the ideal way to do this but what if you have ~1ml left in an LC syringe, could that colonize into 500ml of myc water? Also when using a grain jar for GLC, Agar uses a 60ml syringe with an 18g needle; what if you have a 20ml with a 16g and an 18g may not fit through the lid, will a 16g suffice? And how much water actually needs to be aspirated into the grain jar and then back out of it into the jar for myc water? Agar uses ~160ml however that's about 8 transfers in and 8 out; would you flame sterilize the needle each time you go from one jar to the next? I.E. Flame, Stick needle in jar to get water, take out, Flame, stick needle in grain jar to aspirate water, take out, flame... Rinse repeat? Agar mentions nothing about flaming the needle.
Thank you.
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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ISOfantastic
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Registered: 06/23/11
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Always flame sterilize or wipe your needle down with an iso-soaked paper towel between inoculations of any kind, it prevents cross-contamination.
Honestly, GLCs are misleading, contam-prone, and if you don't have a really good eye for it, it's easy to mistake the white mycelium of trich and other molds for your cubensis mycelium. I'd recommend looking at the link for LC in my sig. Very simple, very very effective.
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Fungus-Amongus
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This is a bummer, I was just about to put water in the jars and PC them. How long is a syringe sterile? Until the plunger is pulled back or the container it is in is cracked open?
Also I assume PC the jars for 30 min and then let cool? How cool?
EDIT: Also most brew stores are closed ATM and I'd really rather complete this task tonight. Grocery stores carry it?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
Edited by Fungus-Amongus (01/14/12 04:33 PM)
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Fungus-Amongus
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Anyone?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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Registered: 04/17/11
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GLC's work well IME. LC's you need to shoot a test jar to make sure its clean with a GLC you know your syringe is clean. I found the grain will suck up 10-30ml of water before I was able to draw any up. This is totally going to vary grower to grower. A larger gauge needle is fine. You should flame sterilize in between each transfer
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Fungus-Amongus
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Awesome thank you, as far as making myc water using a LC syringe?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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Quote:
Fungus-Amongus said: myc water using a LC syringe?
Huh? I know of people injecting elme water into a glc to make it grow some more. What do you mean by myc water and lc syringe?
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Fungus-Amongus
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Turning ~1ml of LC syringe into Myc water?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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undeadninja2
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Registered: 06/15/11
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I think he means knocking up more LC with remaining LC in syringe
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Wing
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To make a GLC you need to get the mycelium off the grains, hence the term Grain Liquid Culture. If you're not working with grains then it's not a GLC.
-------------------- My Old Grow Logs
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Fungus-Amongus
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^^ Yes but with just water, no dextrose or anything like that.
When do syringes become un-sterile? When you break the seal on their container or when you actually move the plunger?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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Wing
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Well you could use dextrose or malt. That would be like a GLC+ LMAO
-------------------- My Old Grow Logs
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UnnamedGrower
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GLC - Water containing myc. This is made by injecting water into a fully colonized grain jar shaking and drawing the water back up. A BRF slurry could be considered a GLC.
LC - Water and some sort of sugar. This is made by sterilizing sugar water and inoculating it. This will grow and can be used to inoculate. I say sugar water but sugar isnt a good option. I personally like ELME, but you can use honey, karo. SITR among other things.
I think the sterility of the syringe isnt broken til the plunger is moved but im not sure
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Fungus-Amongus
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So can you make a "GLC" with a LC syringe? Instead of the traditional way?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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What do you mean by lc syringe? No you cant make a glc from an lc syringe. You make a glc from a fully colonized jar. You need to grow an lc
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Fungus-Amongus
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So the water just basically suspends the myc in a GLC? If I want to turn this 1ml of LC left in the syringe into say 500ml, I suppose I'm going to need dextrose?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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dex isnt a good option. You could either inoculate a jar with whats left and make a glc or make an lc with honey karo elme sitr or something
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Fungus-Amongus
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Hmm I suppose I'll just do another jar with it instead. Are GLCs pretty much fail safe as long as there is flame sterilization while transferring the water to the grain and vice versa?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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Nothing in mycology is fail safe. The best you can get is a flow hood and you still encounter contams with that. If your careful you should be alright
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Fungus-Amongus
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Of course, I know that sterility is key here. Does the Myc actually grow in the water? Or do you need the same amount of myc as water?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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The myc grows in the sugared water
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UnnamedGrower
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There are two links in my sig about teks look at the lc teks
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Fungus-Amongus
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I understand that, I'm more focused on the GLC at this point using Agar's tek. Does the water just suspend the myc or does the myc actually grow in the water? Meaning can you use a tiny amount to fill 500ml of myc water?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
Edited by Fungus-Amongus (01/15/12 01:21 PM)
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UnnamedGrower
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Its suspends the myc. No growth.
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Fungus-Amongus
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Thank you
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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Viruk
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Basically an LiquidCulture is a liquid suspension that contains sugar for growth. This is done by either taking spores (which has a low rate of success IMO)or you can take mycelium from an agar plate, or a piece of a mushroom itself, and place it in the LC to grow more mycelium.
A Grain Liquid Culture, or GLC, is just sterile water that you placed in a jar already colonized with mycelium. You shake this jar either before you put the water in, or after, any way that makes little bits of broken mycelium float in the liquid, then you place it all in more water to expand your GLC. The idea behind this is 1 tiny thread of mycelium can grow an entire new jar of mycelium.
Think fire here, you are just transferring an ember from one place to another.
Keeping it sterile is pretty easy if you keep the syringe sterile.
Basically the syringe is not sterile the second its out of the pack, or the second the plunger is drawn up, or the cap is taken off the needle.
If you draw up the plunger and the caps on, you can still suck in air, syringe is then not sterile. If you touch your syringe to anything, it's not sterile. Once you even inject into a jar, assume it isn't sterile, and burn the needle, wiping with alcohol can also work, but you don't get the inside of it, so lots of people will argue it's not sterile.
In my experience the syringe will stay sterile quite a long time, literally until you break one of the above rules. I've taken a full GLC syringe of 30CC and injected it into 2dozen jars one after the other, without flame sterilizing only wiping, and had nothing contaminate. (since everything was going out, I never pull the plunger up, nothing goes inside the needle) It all depends on how clean you can keep things, if you go insane with your work area you'll be fine, and if you have great technique you'll be better.
I keep LC's to make jars to make GLC's out of, when the GLC runs out I go back to the LC, its a pretty good strategy for keeping things going. You can always make up more jars with the spores you get, and get a new LC out of those.
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Wing
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Re: GLC Questions [Re: Viruk]
#15672813 - 01/16/12 08:59 AM (12 years, 5 months ago) |
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I've had GLC's grow a little more while in the syringe. I've noticed the darker the fluid in the glc, the more likely the myc will expand.
I'll post some pics when I get a chance. The syringe I have currently went from a brownish to an almost white after 4 months. I've just held on to it to observe.
But commonly glc's don't expand much.
-------------------- My Old Grow Logs
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Fungus-Amongus
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Re: GLC Questions [Re: Wing]
#15673297 - 01/16/12 11:17 AM (12 years, 5 months ago) |
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Awesome, good info! Well I have a jar of sterilized water sitting in a ziplock right now waiting until I can get to it (too many people in the house atm). I have made self sealing injection ports with RTV silicone so that's to help towards sterility, on top of that all air will be off in my room of choice and I will turn my box sideways so the top faces me and gives me a surface to work on (knocked up all my jars this way with none contaming).
Syringe container has been cracked open at the store so the lady could see what I needed, plunger has not been pulled back and I don't believe the thing has really even been pulled out of the container. So I'm betting that's safe as well. I'll flame sterilize after I go from the grain jar to the water but it's a 20 ML syringe and that is going to be a lot of back and forths, is it really necessary to flame it each time if each has a self healing injector port and its wiped down with ISO?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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Fungus-Amongus
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Also having a hole the size of a Phillips head covered with Tyvek and another covered with RTV for a self healing injector. These holes aren't too big are they? There's others I read mentioning a pencil sized hole?
-------------------- Anything I say or do on here is all made up and fictitious. Any pictures I may have posted or thoughts and concerns I may have shared are not my own but those of others many years prior.
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UnnamedGrower
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1/4 inch for ge 1/8 for your ship
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