HOW TO MAKE LIQUID CULTURE FOR MUSHROOM GROWING, Non-Vented Lid Jars from Start to Finish
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- Опубліковано 29 жов 2021
- A comprehensive overview of how I make my liquid culture from start to finish. For me, this is the first step in growing your own mushrooms at home. In this video, I focus on using NON-VENTED LID JARS. With non-vented lids, you need to equalize the pressure in your jars manually after sterilizing them in the PC. I demonstrate how to do this step-by-step and discuss the situations in which non-vented lids are preferable over vented lids. Your mycelium will not grow as vigorously with non-vented jars, so you can use a smaller gauge needle in your syringes. I typically use an 18 gauge needle. Smaller gauge needles can save some wear and tear on your injection ports.
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Here's a link to Microppose, the company with the awesome stick-on injection ports and filter discs:
microppose.com/
I think it's beneficial to watch my Liquid Culture 101 video first before watching this. That will help you decide rather a vented or non-vented lid set-up is best for your application. If you're a beginner, you may find the vented set-up more user friendly as there is no need for manual pressure equalization. Here's the link:
ua-cam.com/video/NvGWP_jFZmo/v-deo.html
I think my ultimate favorite part of video two is your extremely effective but simplified way of depressurization/drawing fresh air- I’ve seen other videos which suggest silly techniques like trying to draw air out of the middle of a flame and then push it back in, this simple alcohol air filter technique seems much more reliable and simple!
One more question on LC types- what is your take on recipes which suggest to add a tiny amount of brewers yeast? It seems counterintuitive to add another, much faster growing, organism to the culture- even if you are sterilizing. I understand that yeast is nutritious but the amount is so tiny, I can’t fathom how it would be beneficial enough to be worth it?
@@ChristophHalverson-iw8gk There are tons of recipes and nutrient additives. Brewers yeast has been used for a long time, I wouldn't worry about it hurting your LC, but I like to keep it simple. 4% karo and a pinch of soy peptone, everything loves it.
Great videos! Hey quick tip on the rtv silicone if you spray a teeny bit of soapy water on its surface and your fingertip (gloved if you wish) then you can shape the silicone without it getting stuck to you! lets you make it thicker without having to go wider, if that makes sense. Go on, give it a whirl!
Thank you very much for sharing your expertise.
Much appreciated ❤
Thanks for the fast replies, your 20 yrs experience is a great asset . I appreciate it.
New sub here, and one word: WOWEE! This was so interesting and made so much common sense. The close proximity of your torch with the ISO had me on the edge of my seat, but the meticulous steps you took clearly map out the steps. It was particularly satisfying to watch the LC bubble when the air FAE is injected.
Glad you enjoyed the video 👍 I'm always here if you have questions. Thanks for watching and subbing.
Best mushroom channel on UA-cam. I’ve learned 95% of my mycology knowledge from this channel. And any time I have a question you usually have a video to answer it. When your equalizing the jar with the needle syringe, guessing just putting the needle into the jar for a few seconds, the vacuum inside the jar will pull out the amount of air it needs without any need for a person to manually push the plunger?
That's really nice of you to say, thank you 😁 Yes, it only takes a couple seconds for the pressure to equalize, no plunger needed.
@@RenegadeMushrooms thanks for the reply! Keep up the great work and thanks for sharing all your knowledge!
Thank you! This was awesome!
thank you for this i was searching for how to make a diy injection port
Never new about master jars! Thanks mate
Cheers new to growing mushrooms looking for good info that you have provided thanks mush lovin
I'll refer you to and online store where I got my own psychedelics and microdosing stuff very good reliable vendor
I get my psychedelics and microdosing stuff from an online store where I got my own his on Instagram and also on Telegram with the below handle
Great video. Thank you
Good stuff, good info in your video's
That's slick thank you
Made flo hood work's well glad i watching again soon I'm going to do long term jars now we're going thanks for videos really helpful grow on .
Awesome, having a flowhood is a game changer.
Good tricks. Thx. 🙂
Thank you!
Very nice
We are just starting out and following your videos as gospel! Thank you so much. Just not sure at what temp range we should store them until ready.
Thank you
thats a neat tek with the torch. I'll experiment with that. as for now I can imagine sucking sterile air through the flame inside empty syringes right before drawing out the LC for use. that way I press sterile air indise my jar to get positive pressure while sucking LC back out right after. no more retracting syringes. great video!
Good idea, thanks for watching
thanks man
Nice video. Thank you for sharing your long run of experience. That is so valuable! I have been enlightened about the lack of need for venting the lid during colonization. I've wondered about that.
Two questions - Why vent the LC immediately. It could certainly be stored with vacuum. In theory, would that be best for sterility. Should you accidentally introduce a contaminant during venting, it would be better to do at the time of inoculation rather than give the contaminant a head start.
Second, if you consider the effectiveness of a lami flow filter such as yours and the couple hundred or so ml of air needed to vent the jar, then the chance of a contaminant with by venting with a new or flamed needle should be miniscule. No harm done of course with the IPA syringe tech.
BTW, for those without a flow hood, consider a sterile 0.2 micron Luer lock syringe filter atop your needle as you puncture the injection port to equalize pressure. They are super effective as a micro air filter.
Cheers and thanks again.
Yes, the syringe filter would work well for venting also. Theoretically, an open needle in front of a flowhood would work as well, but I've always preferred multiple levels of security mostly because I'm working in a regular old basement with an old forced air furnace so I have some sketchy air currents. If I was in a clean room with filtered air, I wouldn't sweat it as much. As far as venting a jar right away or leaving it in vacuum, you could leave it in vacuum until you're ready to use it. But, if mold or bacteria sneaks in there during venting at any point, your jar is toast. The advantage to venting them immediately and then storing them for later use is that any contam will germinate and you'll know not to use that jar. I've vented them immediately, and stored them for months before inoculation with no issues using this method.
Thanks for the response. I have watched several of your videos and learned a lot. I was thinking about a hybrid system that has the advantages of the vented lids (no need to equalize the pressure with a sterilized syringe after the PC, etc.) , but also the advantages with non-vented, ( i.e. no thick mycelium, cultures last longer)
I find both are useful, so it's good to know how to make vented and non-vented LC.
I was thinking you could use a syringe filter to equalize, then use a syringe cap on the filter to stop the f.a.e. disadvantage to this is you are not eliminating any time or cost on the lids, and stacking sucks with syringe filters. Let me know if you came up with any good ideas!
I was looking at my vented Pink Oyster, so chunky thought it had developed a spine.
Gracias👍
thankQ 4this!
So informative, thanks for dropping the knowledge. once i slow down and learn its so easy. So micropose, i love them! Question, can you put an injection port on both sides or is that overkill?
Two injection ports isn't really necessary. I always look at it like any additional holes in your lids are potential contamination points.
@@RenegadeMushrooms i chill ville means like 2 on the same hole 1 on top and one on the inside, the same way the rtv is done but with the self adhesive ports
@@angel2jimenez219 Not necessary.
I have learned a lot from your presentation. I always thank you for your presentation. I have a question, after making cordyceps liquid culture, how long can I keep and use it?
You want to use it within 2 weeks for best results. If you need to store it longer, it's best to keep it in the refrigerator. A fresh high quality culture will store much better than an older weaker one, but, in general, cordyceps cultures senesce much more rapidly then other species. So, the longer you store them, the weaker they get.
The. MASTER 👍
@renegade mushrooms
I like what you said about not needing to keep the liquid cultures in complete darkness. thank you for that. I was wondering is this also true for temperature? Does that also not really matter or is there a temperature range that is really ideal for keeping newly formed liquid cultures? Thanks a lot I love your channel.
Glad you enjoy the channel 👍 My basement ranges from 60 to 70 F depending on time of year and I see little, if any, effect on the growth of my LC in that temperature range. I would say anywhere from 50 to 80 F is fine.
@@RenegadeMushrooms thank you for taking the time to answer my question
liked and subscribed
Great video, I've been learning lots! I did have a question after the point you take the plunger out. Was the entire process just to clean the potential 'bad air' in the needle and after that, because you are in front the the laminar flow hood there is less (no?) fear of contamination when the jars are equalized with the air in front of the hood?
Yes, it is to clean any potential contaminants out of the syringe and needle.
Awesome video!
(beginner here)
I thought of a solution that might work:
what if you take a syringe needle, you put a micropore tape on the needle large opening (the needle-syringe connection side), you pressure cook the jar with the needle in the injection port (to allow gas exchange) and when the cooking is done you remove the needle.
Do you think it could work?
Yeah, it might. I actually really like that idea although I would probably go 2 or 3 layers of micropore to be safe. Or maybe glue a small filter disc to it. I'm gonna try it 👍
@@RenegadeMushrooms Awesome, let me know if it works!
@@RenegadeMushrooms
Can one use plastic jars instead of glass jar?
@@bestheirs8999 Plastic jars are ok as long as you use thick PP5 plastic, but glass is superior in my opinion.
(Currently just a spectator in the depths of a monthslong binge on this subject, excuse my ignorance.)
Would adding some amount of H2O2 suffice to “vent” (or essentially add back both oxygen and volume to avoid hypoxia and a vacuum) rather than this intense process you’ve got here? I imagine since you’ve been doing this forever you have some reason(s) why this isn’t done. Thanks for all your work man, this content is fantastic.
That's an interesting idea, but you would have to add 50 ccs or so of H202 just to equalize a pint jar. I'm thinking that would cause issues with growth. You would need a monster syringe, or have to do multiple injections. This air venting method might seem complicated in the video, but it's actually pretty easy once you do it a couple of times. Thanks for watching 👍
Thanks again for the video. I want to use the equalizing method you demonstrate in this video but don't have a laminar flow hood. If I do it inside a SAB, could I then wait to see if anything else starts growing in the jars before innoculating them or would you recommend innoculating right away. If OK to wait, any idea how long I should wait before expecting to see contaminates growing?
That method will work fine in a SAB, just be careful with the flame and alcohol vapor. Flame outside the SAB, then go in and equalize. I would inoculate right away if it were me. Just always test a bit on a small grain jar before going big time.
What is the best way to make a flow hood can i get a plain somewhere thanks for your videos all good know how I'm just getting going with it
There are lots of designs out there. I prefer a centrifugal fan setup myself. I explain how I built my hood in this video:
ua-cam.com/video/AE9tP61JgP8/v-deo.html
Yeah, love your videos! Question: LC and stuff all works perfect and now i wanna store some like you are describing. Can I not just use a vented lid and cover it with scotch tape whenever, and when drawing just remove the scotch tape? I know there's a lot of things that work - but would love to know what you think or thought about it. Thank you 🙏
You could try it, but my concern would be that the filter patch would grow mold under the scotch tape. The filter patch needs to exchange air and do it's thing to keep it dry. If you cover it with tape, the excess moisture build-up from lack of airflow may cause mold to germinate on it contaminating your jar.
Have you tried Rutand RTV 500° F Clear Silicone for the self-healing injection ports? The benefit is that the drilled hole below can be seen otherwise it has the same max temp as high temp red gasket RTV silicone.
No, I never have, but I'm sure it would work. I've never really had a problem not hitting the hole though with the red.
Great video! I was wondering if you had one on agar? I'm having a lot of contamination and wonder what you do. ☺️
I've used liquid culture for many years, but I just started working with agar and I'm enjoying it as well. I really only plan to use agar to clone wild mushrooms. I recently made a video of cloning a wild Morel on agar if you are interested. The project is on-going.
ua-cam.com/video/G2SiinsEcCI/v-deo.html
@@RenegadeMushrooms Great thanks!
Could you simplify this by having a waterfilled jar with filter among theese with only injectionport and take air from the decomressed jar through the injectionport? Just follow the same procedure with isopropanol sterilization but without open flame.
Yes, that would probably work, but it may be more work than it's worth as it takes a lot of air, usually more than one syringe, to equalize you jar pressure. Here is a link to a new technique I just tried that works great:
ua-cam.com/video/28uUicG6M2g/v-deo.html
Awesome video and learning about this stuff. I do need to ask why you are making so much LC? You only use 2cc to inoculate grain jars, right. By my math, you could inculcate 1200 quarts😀 Just in case one or two jars get contaminated? Still a ton. Thanks.
I'm doing multiple different species of mushrooms, hence the multiple jars. Also, sometimes I use a whole jar per grow with the mycelium flood method I demonstrate in my video on how to grow Cordyceps.
Thanks for sharing your experience! I have a couple questions and forgive my ignorance it’s been a long time since I’ve had any science classes-
I don’t understand how the air stays sterile after the plunger comes out. Why not use the plunger to push the enclosed sterile air into the jar?
Would it be possible to seal the vent hole after sterilization without contaminating the jar?
There are a lot of ways to accomplish the pressure equalization step, this is just what has worked for me for 20 years so I feel comfortable that it will work for others consistently.
The air in the syringe stays sterile because you are flaming the needle as you pull back the plunger. It takes a lot of air to equalize the pressure, so if you left the plunger in it would take 4 or 5 syringes worth to equalize one jar. This process has redundant steps to ensure clean air enters the jar.
Hey boss, were can I get empty syringes? You order online? Thank man. Keep up the great vids
Thank you👍 I ordered a 30 year supply 15 years ago from a medical supplier in Jersey, but they seem more difficult to find nowadays. Mycology suppliers seem to be the best bet now. Check out Shroom Supply or other online mycology stores.
Great video man!! ?? Where do I find the 12 ga needle? I have looked every where. Can you send link please.
I've had trouble finding them myself lately, but I know Shroom Supply has 14 gauge needles and they work well.
Ty so much! What micron is micropose? The reason for the poly fill syringe?
Sorry for the late reply, I missed your comment. The microppose filter discs are .22 micron. The polyfil syringe process I demonstrate is the technique I use to manually equalize the pressure in liquid culture jars with no filter disc in the lid after running them in the PC.
When you pressure cook the jars do you tighten the lids? Also do you turn the lids upside down? Thanks I subbed
I snug the lids down tight and they are the right way, not upside down. I've never had one explode on me, but I only fill them to 300 ml in a pint. If the jars were full of liquid, they might break on you.
13:10 i like to put a thin folded strip of aluminum foil under the lid seal before the pc and simply pull it out after it cools and I've never had an infection problem. Especially when you put a pressed cotton ball over the vent of the pc for extra clean air pullback haha
That way you just gotta pull the tab without even lifting the foil covers if it's sticking out enough and then simply tighten and boom done. Saves me lots of time with large batches!
Good tip, thank you 👍
Could you put the poly fill syringe in the pressure cooker with the jars to sterilize it instead of the alcohol and torch method? Wrap it in foil or autoclave bag?
Yes that would work too. I've done that before with the foil wrap and it worked well.
Great video, thank you. Question please. You are allowing clean air ( a few CC's ?) into a LC jar that is under vacuum to equalize the pressure in the jar to match the outside atmosphere. Then you inject about 10CC of LC back into the jar causing a positive pressure situation in the jar.
Would it not make sense to just skip the equalization step because you are going to add 10 CC of fluid into the jar, there by equalizing the pressure that way. ?? Thanks
It's more like 200+ ccs to equalize.
@@RenegadeMushrooms Wow, thats a lot of vaccum ! I wasnt aware it was so much. I understand more clearly now. Thank you for the quick reply. I enjoy your videos so much, I re-watch them every time I go to perform some mycology steps. I'm still a noob. Oh, I tried your trick about injecting sterile water into a colonized grain bag and then aspirating the water back out and then injecting it into a fresh grain bag, It worked!!, no contamination at all. Thank you again :)
@@lycanzhp1141 Awesome, glad it worked for you 👍
Thanks for sharing your knowledge. I was wondering if I can replace polyfil with cotton?
Cotton, being a natural fiber, absorbs water and is more prone to contamination. Polyfil is cheap and definitely the way to go.
@@RenegadeMushrooms Thanks for the quick response. I was asking this because polyfil is hard to find near me, only online. I also understand that reuse is the way to go. However, as I don't have polyfil, I'm going to do a test with cotton, but I'm not going to reuse it. Thank you, I learn a lot from you
@@crypto.entrepreneurs You can also use 2 layers of micropore or cloth medical tape if you can find that.
@@RenegadeMushrooms Yes that I have. I'll try it that way too. Thanks
Hey Renegade…..can you use spores to colonize your liquid culture? Love your vids!
You can as long as the spore syringe is sterile, it works great. Thanks for watchin 👍
@@RenegadeMushrooms hey Renegade, I subscribe to over a hundred channels covering various subjects. You are the only one that I know will answer a question. Great job! Really appreciate it.
Hello, I'm in an area that used 10 psi, should i still use 15 psi? What is the purpose of the tinfoil tops,? Would an 18g needle be ok to use? Thank you, i love your videos :)
If you can only do 10 psi, I would go for 60 minutes instead of 30 minutes. The tin foil tops are to protect your injection ports and/or filter discs from steam in the pressure cooker. 18 gauge needles are the smallest you can go. They will work, but you will likely get some clogs. 14 gauge works pretty well.
I am wondering at what temperature do you keep this culture at after until it is ready to use? We are keeping the cultures I made in a wine frig at 50-60 degrees
A wine fridge is a great choice actually for LC storage of cold tolerant species. Just remember that tropical species like pink oyster and almond agaricus should be stored at room temperature, as temps down in the 40s can harm or kill them, 65F is good for them.
Hi, interested in trying the step to let sterile air in, do you mind sharing a link to the needle used. I understand it’s heavy duty 12 bf needle? TIA
I'm actually using a 16 gauge BD needle that I got for free with a LC syringe.
When you reuse your polyfilled syringe. Do you just refill it with iso and flame it, or do you need to run it through the pc as a precautionary measure?
Just refill it with iso, push through to clear all the iso, then withdraw plunger while flaming. I've never run it through the PC. Works every time for me.
So while the jars are colonizing should you shake them at all?
I assume you would definitely shake them once your ready to draw a syringe full. Sorry if it’s a dumb question I am brand new to this.
Not a dumb question at all. It is a good idea to give it a swirl or shake every few days during colonization. Just make sure, if you choose to use a filter in the jar lid, that you aren't continuously getting it wet when you shake because it will eventually grow mold. If you use a lid without a filter, you can shake at will.
Thanks for the cool video. Never heard before that lc doesn’t need air exchange. One question. Couldn’t you just use vented lids and keep them covered with aluminum foil really snugly after you take them out of the pc? You only had to remove the foil when you inoculate or draw from the lc. I guess that wouldn’t completely stop the air flow but limit it enough to get some of the non vented lid benefits.
But after all you put aluminum foil over the jar lid anyways once they go into the fridge for storage.
The main drawback to having vents in the lids is the vents are prone to have mold germinate on them if you don't keep them dry. So, if you're not extremely careful when pulling syringes, you can continually soak them with LC and covering with foil can actually hurt in that case because it will cause the filter to remain moist. Once mold germinates on your filter it will punch right through to your jar and its game over, so you really have to focus on keeping those filters dry if you're going to use them. For this reason, I only use filters on jars where I'm going to dump the whole jar to substrate like with my mycelium flood method for Cordyceps and Enoki.
@@RenegadeMushrooms Hello Thx you very much for the quick reply. Yes. Actually I just started growing shrooms. I used 2 layers of micropore tape for venting , but those are constantly wet. I guess because the liquid is breathing through them. Non vented lids are probably a great alternative.
I also used spore solution to inoculate the liquid culture (as recommended in a certain book). Problem: The spores multiply but don’t germinate. Lc works much better to inoculate lc. 😉
Could you just use a vented lid, then after the jars cooled and pressure equilibrated, seal the filter disk with tape or RTV ?
If you were going to do that, then you would just use a lid with a filter disc in it like I show in my 101 video:
ua-cam.com/video/NvGWP_jFZmo/v-deo.html
Hi! would the nooks and crannies on the red silicone injection ports make it difficult for the alcohol to do its thing? The stick on ports seem less hospitable to unwanted guests
No, it's never been an issue for me but when you apply your RTV you do want to make it as smooth as you can. Less nooks and crannies vs. more.
@@RenegadeMushroomsFor sure! You have a ton of videos and its clear you enjoy making these...
It would be awesome if you put out a blooper reel one of these days
More questions for you...why couldn't you just sterilize the needle without the plunger attached? Is it because you wouldn't be sterilizing the needle all the way up to the plastic fitting? Also, since you're still introducing the air that's in front of your hood, wouldn't it be nearly the same risk as if you just cracked the lid briefly or no? Also, how would you do this with a still air box given the use of alcohol and flame? Thanks again.
Yes you run the alcohol through first to sanitize the entire pathway of the incoming air, needle hub, syringe, etc. It is not the same as just cracking the lid in front of the flowhood because you get an extra level of protection with the syringe because all of the air flows through the alcohol sanitized pathway of the syringe, polyfil, needle etc. Just a more controlled way of doing it. Everyone loves to think of ways around doing this, but it's always worked for me so I've stuck with it. I really don't find it difficult, but it's not something I would attempt in a SAB. If you are using a SAB, just use a filter disc in your lid in addition to the injection port and problem solved.
Will that work with a syringe full of spores? Inoculate a jar with a syringe full of spores?
Yes it works well with spores too as long as the spore syringe is clean and sterile.
Do you think if i make it with Tyndalization instead of pressure cooker will it be fine?
That's a very interesting idea. Theoretically that should work. The only issue I could see is that it may lead to excessive caramelization of the sugars in your solution, but that may not be an issue. Try it and let me know if it works 👍
Put an extra jar with a filter & 50ml water in the pc, draw from that one to equalize the non filtered jars, I have flat .2 micron paper filters for this & you dont even need to pc it again, everytime you pull with the syringe air is filtered on its way into the jar.
Cool trick 👍 This alcohol soaked polyfil deal is pretty quick and effective as well.
Would you use this method for a bigger jar? Or would the larger jars require more fresh air?
It works fine for quarts, never tried it for half gallons. I'm sure there is a point where the liquid volume would require a filter to myceliate sufficiently.
Hey, I had a quick question. Would having a sterile syringe & needle right out of the package eliminate the need for the polyfil and flame? I would imagine so, just curious if you've had a bad experience with it.
Also, great video! Thanks for the helpful information!
I've never trusted just sucking in air from in front of my flowhood right in to my LC jars. The alcohol soaked polyfil serves as an insurance policy to catch any contams that may be present. It's never failed me in 20 years, so I stick with it. A needle right out of the package would not need to be flamed. Thanks for watching 👍
@@RenegadeMushroomsaway another question, please: why couldn't you tape over a vented lid filter with regular tape, to prevent further air exchange?
@@stephenwalther9702 I don't see any reason why you would want to do that. The only reason you wouldn't want a filter in the lid is because then you don't have to worry about getting it wet when you shake your jar, potentially causing mold germination. Just taping over it won't prevent that.
what size are those black self-healing plugs please? I just searched for them and found they come in two sizes.
microppose.com/products/adherable-injection-ports-for-plant-mushroom-tissue-culture
Don't forget you can use promo code "RENEGADE" at checkout to save yourself 10%.
Do you think the pickle jar lid and a hole with medical tape would work?
Probably as long as you incubate in a storage tote. I would go with two layers of medical tape though. Also I wouldn't use a jar bigger than a quart with just a single hole. Anything one quart or less should be fine for air exchange.
A (kinda obvious, but i forgot about it aswell) point about the RTV silicone ports...seems to work fine with his lids and the big diameter hole but mine were plastic (PF jars) with a hole half the size...should have put some sand paper to the bit around the hole...the Silicone didnt stick to the flat plastic at all and is just held in place by the silicone in the hole.
I've never had a problem with the high temp RTV sticking to the metal lids, but I can see that being an issue on plastic if you don't rough it up a bit.. Thank you for sharing your experience.
Could one stuff a synthetic filler disk in the syringe instead?
You could use a syringe filter, but you need to make sure everything downstream of the filter is clean as well. There are lots of ways to vent jars, this is just what I prefer, never let me down.
Would a stopcock work for equalizing the pressure?
Yes, if it was mounted to the lid, and you used some type of sterile filter media.
Do you reuse the foil lids? I never thought about that.
Yes, I re-use them many times, until they get brittle.
Hi, Can I inject the liquid mycelium straight onto wood or straw.
You can, but you need to use a lot of LC if you're going to spawn it to bulk substrate. It will be slow to colonize as well, so it's not very efficient and leads to increased risk of contamination. That's why the general rule is LC to Grain to Bulk Substrate.
Can spore syringes be used with method?
Yes, as long as the syringe is sterile.
@Renegade Mushrooms: Hello. New to growing. Am I wrong to think I could affix a 2u filter between a needle and plunger-less syringe and then use it to equalize pressure? Serious inquiry.
Yes, as long as you assemble everything in front of the flowhood, that should work fine. The alcohol soaked polyfil is an old school poor man's trick from back in the Shroomery days. It's always worked flawlessly for me so I stuck with it. Try the filter deal and please let me know how it works for you.
@@RenegadeMushrooms Thank you! I will.
Do you need a torch or could you do this with a lighter?
You can do it with a lighter, but a small torch works much better. You don't need a giant propane torch like I have. A small butane torch is fine.
Can I buy syringes in local stores or do I need to order them online?
Better to buy them online imo. This is where I get mine, they are an affiliate of ours:
www.shroomsupply.com/syringes-needles/sterile-syringes/?referrer=CNWR_684691708647669
How do you pressure sterilize with an instant pot?
I've never used an Instant Pot so I don't know. I'm sure you can find some vids on YT.
oh yeah I see a lot of videos. I didn't know it was even used but I guess some people prefer it. I am going to try to grow lions mane for my first mushroom. any tips?
@@RenegadeMushrooms
@@AM2PMReviews
Lions Mane vid:
ua-cam.com/video/N-Qpb3K8EL0/v-deo.html
I think some people prefer the instant pot because it's cheaper than a real pressure cooker. If you're going to get somewhat serious about this hobby however, spending a couple hundred bucks on a 23 quart Presto pressure canner is well worth it imo.
Wouldn't just squirting (the jar actually sucks) the LC syringe equalize the pressure inside the jar ?
Not even close, you could dump a full 20cc syringe in and still have vacuum.
question, why couldn't you use vented jars for sterilization and just replace the tape with non porous tap under the hood? to slow the growth down?
You could, but when you're keeping a jar around for a year or so, and pulling dozens of syringes from it, there's a significant risk that a taped hole becomes a contamination vector at some point. Not worth the risk imo.
@@RenegadeMushrooms yes every weak link... I'm small time just starting out, looking for ways to prolong liquid culture if I get that far
I love this shit
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Correct me if I’m wrong but couldn’t you just sterilize the culture with foil over the top, open your cooker in front of flow hood, cap your jars then inoculate? This should eliminate the vacuum problem.
I imagine the temperatures would be so hot that any steam/water from the cooker that mixes in will then be sterilized in the process and be free of contaminants, have you tried that before?
There are a lot of potential contam vectors in your proposed process unless you're working in a very controlled contam free environment, which I definitely am not. Foil is a very imperfect seal, removing foil and capping could introduce contams, and a cooling PC can pull in contams, potentially introducing them to imperfectly sealed jars inside. Their are lots of ways to successfully sterilize LC. You can also put a filter in the lid that will automatically equalize pressure without introducing contams (I show that in my other LC video). All I can tell you is that the techniques I demonstrate have worked flawlessly for me for 20 years, but there is always room for improvement 👍
So, I've now made two nutrient jars for LC, pressure cooked them at 15psi for a half hour, and both times, about 2/3 of the liquid evaporated from the jar. Today, that happened, and when I removed the jar, the bottom cracked off! It was a real one pint Ball canning jar, too... I let it cool off in the cooker, but it cracked after I lifted it out. Don't understand what's going on.
I'm not sure either. What lids are you using? How full are your LC jars with liquid, and is there water left in the bottom of your PC when the cook cycle finishes? Also, what are you using for a heat source? I've run thousands of jars, and never had one break.
Using a pint jar and your exact proportions, do you have any idea what volume of air would be needed to equalize a jar? If you were to use a large cc syringe could you not remove the plunger when equalizing? That way any air entering the jar would have flowed through the hot needle? What do you think?
Also can you suggest location to purchase 12/14 gauge needles? An internet search isn't bring up much larger than 18 gauge.
I've tried it with up to a 50 cc syringe and it wasn't enough. Maybe 100cc would work? 14 or 16 gauge needle is fine, 12 is a little big so I need to edit that comment. Shroom Supply has 14 and 16 I believe.
Hey RM, is there any disadvantage of using the good plastic lid (with the appropriate silicone seal) when making non vented lids (using the red RTV)? Or do you recommend metal lids? Thanks!
@@mse1333 If you're going to go with plastic lids, I highly recommend the Masontops Tough Tops lids. I wouldn't trust any other plastic lid set-up I've tried for LC. Or just go with the old school metal canning lids.
Plus I the wild, millions of spores are produced with only a small fraction of them ever colonizing and producing fruiting bodies for us to enjoy.
Are the RTV lids dishwasher safe?
Yes I would think so, but I always hand wash them, only take a second.
Hey RM, so why do you only fill the pint jars to 300ml, any reason? Also if you were to fill the jars almost completely, there would be no air so little to no vacuum would be created during the PC processing, so maybe no need to deal with equilibrating the pressure?
There would still be enough vacuum that you would need to equalize. You could put a little more liquid in there if you wanted, but overfilling them can cause alot of issues, so if you need more volume, just make more jars.
If the jars were filled almost completely, would there be enough oxygen for a year's storage?
@@RobBanks81 It's possible they may be starved for oxygen if the jar was too full. It could cause all kinds of problems actually, no reason to do it. I've always filled them to 300 ml. for a pint and 600 ml. for a quart and never had any issues.
So even 18 gauge needles are not sufficient for the thick myc from a vented lid? 😳 I can't find any smaller gauges than 18 where I live
18 gauge works fine if you don't add the peptone to the LC recipe. It will grow fine with just the karo, just more slowly and not as thick.
Where did you manage to find a 12 Gauge syringe tip? Im not having a lot of luck.
They're hard to find nowadays. Shroom Supply has 14 gauge needles the work great though.
@@RenegadeMushrooms I wonder why this is the case. Is preferred amongst drug abusers? 🤔
Obviously this is working for you but I have a question, after relieving the vacuum and inoculation the jar it's then under pressure. Should you equalize again?
Not necessary, all you need is that initial equalization to relieve the vacuum. Just allow it to colonize for 2 weeks or so and it's ready to use for up to a year with no additional pressure equalization needed.
@@RenegadeMushrooms
Thanks for the follow-up. One more question, have you ever tried cooking your grain in agar befor inoc to speed up spawn?
@@diverwa No I haven't, but let me know how it works if you try it.
Question: Couldn't you just leave the lids slightly ajar in the pressure cooker with the foil over the top of them and then just seal them in front of a flow hood or clean room?
Technically yes, as long as you seal them in front of a flowhood. The problem is that your pressure cooker pulls in outside air as it cools, so you are risking contams entering your jars in the PC if your lids aren't sealed, although the foil would help prevent that. I work in a dusty old basement, so even with a flowhood I prefer to leave my lids sealed. This method has always worked flawlessly for me and others, but there are many ways to accomplish the same goals in mushroom growing so I always encourage everyone to do what works best for them.
Thank you for your answer@@RenegadeMushrooms I'll have to experiment then and see if there's a more reliable than not way of this method.
Questions please and if you could help it would be amazing thank you. Time is of the essence per se lol.
1. You said no direct light, however would using my flashlight to inspect everything in my lab thoroughly cause any issues or retardation in growth.
2. Can the GS, LC and Agar all be stored at the same temperature or does one prefer different temps during different stages. If so what temps per stage.
3. Using 18g needles, I'm getting pieces of my silicone in the...well everything, the needle cuts out a tiny piece. I pulled an Agar specimen out to inspect it and noticed FAE, so I looked a little closer and there's a hole clean through my S.H.I.P, I generally make them atleast ¼" on both sides of the mason lid. Any advice?
I'm currently using 18 gauge needles as well. All injection ports will wear out eventually, but a good dollop of high temp RTV is the most durable in my experience. I would say I get about 50 injection/extractions out of mine before scrapping. If yours are wearing out prematurely, make sure you're using high temp RTV and putting it on thick.
Your flashlight comment cracked me up because I do the exact same thing and no, a brief exposure doesn't seem to hurt them. Also, I store everything at ambient basement temp which is 60 to 70 F depending on the season.
@@RenegadeMushrooms thank you so every stage likes the same temps. Also I'm using high temp RTV Silicone and it's been the first time I put the needle in. Same with the S.H.I.Ps on some of my plastic lids. First poke and I can visually see the hole. Also it's not that brief of a light show for my babies. I went full production from the start knowing I should start small. I've watched hundreds of hours of videos and read 3 books 2 by Stamets and the mushroom grow Bible. I felt confident enough to dump a handful of cash down on everything I needed including a LFH. I have 25 56qt monotubs. So when I go into the lab I turn the lights on and spend about an hour maybe more inspecting agar cups, LCs and GS. So I'm in there for a few I'd that too long or should I adjust my tek. And a question I forgot to ask... I notice you like to shake up your GS allot earlier than most growers, I understand all the principals involved as well, however, what is too much shaking, when is too early and when is too late. I love your channel and I'm very familiar with good ol awkward Gary. Thanks for your quick response. Also my light is an Olight Warrior 3S so it can go all the way down to half a candlepower and run 1watt. So that should be ok? I was thinking of getting an Ultra Violet flashlight and green laser for some of the inspections, what do you think of this method? Last thing brochacho, I've been manufacturing a synthetic dung via nutrient supplementation, I use it as a nutrient broth when gaining field capacity in my Bulk Sub and it works great! Any time I notice contamination on my agar plates, I drop a few drops of Colloidal silver directly on to the contam, it's worked for everything but yeast and the mycelium doesn't care much and bounces back very quickly. I had a contaminated plate I soaked in Colloidal Silver and then put it up and actually forgot about it for almost a month or more. I stuck my spore syringe into the grain spawn just cuz and it sucked the last of my solution out so fast it hurts. I thought I'd lost my genetics, but I just cut 8 clean plates from that contaminated and treated with CS plate. So I saved the genetics with colloidal silver. I highly recommend CS over H²O² or anything else for that matter. I am going to perform more experiments with it soon lol. Sorry for the novel
@@treehouseconstituents6402 That's a cool idea with the silver. I know it's a potent antibiotic, but never heard of anyone using it for mushroom cultivation 👍 I don't think that brief exposure to a low wattage flashlight would hurt your grows. If you held it on one spot for like 20 minutes maybe, but not just glancing around checking grows. Not sure what's going on with your injection ports, but you may just be over-analyzing. You will always be able to see where the needle penetrated, but there shouldn't be a hole that would allow contams in. If there is, your RTV isn't curing properly or something. I've tried a lot of pre-fabbed rubber injection ports and they all pretty much suck. My favorites are the self-adhesive ones from Microppose.com, but you can't beat RTV for durability.
@@RenegadeMushrooms thank you, and I could see light through the hole. My assumption is that most of the times my needle took a "core sample" lol the core was extricated. So. I just fully processed 32 grain spawn today I have to go to bed it's 5am lmao I didn't even realize. Can't stop mid project though fuck. I can't leave my FC grain out all night all I can do is jar it FML
I'm new to this ,but was wondering if this would work for releasing vacuum in LC Jars. Before putting them in the PC put a old needle through the injection port and cover with tin foil as usual. When completely cool take out of PC , crank the lid down tight pull the needle out of injection port , put tin foil back over lid. Seems like this would keep the pressure equalized and sterilize the whole works including the needle when in PC. Let me know if I'm way out in left field on this Idea.
I actually had a video at one point showing that exact technique, as another viewer had suggested it. While it seemed to work for me, I had several reports of the needle leaving a permanent hole in people's injection ports after the PC run. As a result, I took the video down. If you don't want to do the alcohol syringe trick, you can also use a sterile needle with a sterile syringe filter firmly attached to vent your jars. I understand the alcohol/flame business can be intimidating, especially for beginners.
@@RenegadeMushrooms When you tried it what type of injection port did you use. You seem to use a good amount of RTV I've noticed in a lot of videos they didn't use very much maybe that was the problem. Have you tried it with the stick on ports or the rubber ones that you push through the hole? Well anyway nothing is ever simple, but sometime I might try it with all three type of ports and see what happens.
@@samgodfrey8141 I use a good gob of high temp RTV on my LC jars and it seemed to work fine for me. I was also wondering if maybe some people are using regular black RTV, which may create the issue.
@@RenegadeMushrooms I had that exact issue using rtv, permanent hole. Luckily the rtv was nearby and I was able to put a glob on quickly and saved the jars, but I’m not sure of a great solution. I don’t have a flow hood so now I’m a bit worried about trace air being sucked in when I’m taking out syringes full and creating a new vacuum…
@@goodelleric You might be better off just having a filter disc in your lid until you have a flowhood setup.
Hey this question doesn’t have to do with this video but I figured I’d ask you cause you know your mushrooms!! My question is when you’re making agar cup recipes (if you do that) what could be used as a replacement for a media bottle? Those bottles are wicked expensive. Let me know if that’s something you do. Thanks man.
I've worked with agar, but never poured my own. Pretty much everything I do is LC. Hopefully another viewer will chime in. You could always ask the people at Shroom Supply too, they might have a suggestion.
If you have have a pc then do no pour agar, its what I do because I use a SAB
Hey Harold.... FWIW I've been using the bottles that Kombucha is typically sold in and the grocery. They are about 500ml, clear and typically built like a tank. Their stickers/labels are a bit of a pain to remove so I steam them for a few minutes, peal the (usually clear) wrapper. To get the glue residue off I wipe with something like Goof Off (acetone) or mineral spirits/lantern fuel/kerosene Never had an issue and I figure I've saved myself enough for some great MSS'. Needless to say, you might want to do this away from your lit torch. :)
Question:
So once jar is inoculated does it have to be stirred daily? Or can the mycelium be left alone to do it’s thing?
It will grow if left alone, but it grows faster and won't be so thick that it clogs your needle if you swirl/shake it every day or two. Some people like to put a small marble or stainless steel ball bearing in the jar to help break the mycelium up when you shake. Magnetic stir plates are relatively inexpensive too, and they work great.
@@RenegadeMushrooms awesome thank you!
How long can you keep a master jar going?
1 year or a bit more under refrigeration.
I torched a old needle and just for fun poked it in the cover of a plastic petri dish. To my surprise it went right through like butter , leaving a perfect hole the size of the needle. This got me to thinking why couldn't
I draw a syringe of new LC and violently inject into the petri dish to break up the mycelium and then suck it back into the syringe and inject it back into the jar of LC to inoculate it. Pick this idea apart and let me know if you think it would work.
Sounds solid to me, you should try it. I would melt the hole near the edge of the plate and make sure it's sealed really well with parafilm. That will allow you to tip the plate on edge and make it easier to pool the LC and suck it back out. Let me know how it works 👍
Is the purpose of doing this just so you don't have to keep buying liquid culture? If I were just getting started and wanted to grow one kind of mushroom, would I just buy some liquid culture and keep repeating this process whenever more is needed? Thanks
Yep you got it. With this method you can expand one LC syringe into pints, quarts, even gallons of liquid culture to inoculate your spawn with. Much cheaper than continually buying syringes, at least for species you think you are going to grow a lot of.
I’ve had a bit of bad luck with my LC coming out of the PC looking like a dark burnt mess, any tips?
How long are you cooking it in the PC? It only needs 30 minutes at 15 psi. Usually really dark color is cause by sugar caramelization from overcooking.
@@RenegadeMushrooms I’ve done it a few times and probably cooked it a bit too long, but I was also thinking perhaps I didn’t have it mixed well enough. It comes out all settled to the bottom and clearly burnt
@@ChristophHalverson-iw8gk What recipe are you using, mine?
Is 15 psi due to your elevation? Or just extra sterilization steps? Because I pressure can food and I'm at 600 feet above sea level and it calls for 10 PSI for about the same amount of time. I'm actually binging your videos right now, because I have done some decent mushroom grows, but I want to get more yield for less purchase.
Cool, let me know if you have any other questions. My PC rockers are set for 15 psi, and that's fairly standard for most mushroom growing sterilization procedures. I'm not an experienced canner myself, but I'm sure 15 psi is overkill in some situations. I was taught to run at 15 psi, so that's all I've ever done, and I rarely get contams so I've just stuck with it.
In two and a half hours of time, I have five pages of notes! I live in upper Michigan and I thought the coolest thing (besides all your content) was resting the grains. "So I'm going to go ice fishing for 3-4 hours"
@@grizzlybeast5 Ahh cool, yes fishing is my other passion. I fish all year, but I love to ice fish. I have a good friend in the U.P. and he always brags to me about their long ice season up there. In Western NY, we're lucky to get 2 good months of ice, but I hit it hard when we have it.
LMFAO! He can brag all he wants, but if you have to add an extension to your power auger, ice fishing isn't as fun!
@@grizzlybeast5 Agreed lol
Can you not just suck the air back in infront of the flow hood to get sterile air?
Theoretically yes, but I've always been a fan of redundant safety systems. Even the best HEPAS don't catch EVERY contam and one bacterial endospore or trich spore in your LC and you're boned. I'm also a bit paranoid because my lab is a dusty old basement 😁
Minor point: Heat sterilizes, alcohol sanitizes.
Wiping a heat sterilized needle with alcohol downgrades it to sanitized. This is good enough in this situation.
Valid point. I like to pressure cook a jar of syringes with my liquid culture
That may be, but will the carbon buildup on the needle hinder the sealing of the injection port? Or does the use of a propane torch not cause carbon buildup on the needle? And the idea also was stated to cool the needle so the injection port does not get accidentally melted...
Do u have to use the soy peptone?
No, it's a nice supplement. But just karo will work.
Could you achieve the same thing by having the solution filled jar in a sealed vented grow bag, have the lid loose and after it cools tighten the lid? I still don’t get why your jars don’t explode. If I did the same thing with a tin can where the only hole was RTV, would that explode? RTV would pop out from the pressure?
Yes, your grow bag idea would work. Not sure why the jars don't explode, but they never have. They must be able to vent the excess pressure even when snugged down. You would just have to try the can and see what happens.
Do you happen to know is liquid cultures ever exist in nature?
If the can explodes I have one more angle. What likely would happen if you put a small fresh ball of RTV wrapped snug in plastic and tinfoil, to prevent it from drying, into PC? Maybe try that outdoors. I realize on my new experiment putting LC into a grow bag, I could have added the RTV after PC. PC likely doesn’t stres it cause it’s made for higher temp.
And last, what about PCin Non bpa lined cans, no toxic fume melting plastic concerns? I have a lot of questions but I want my experiments to be safe. Peace and thanks again. This vid inspired me somehow to create 2 cool Tek. And now I see I can keep my LC in test tubes, don’t want my roommates to accidentally drink LC in unopened looking soda cans.
EUREKA
But put it in a grow bag with the filter right near the top where you heat seal it in. PC, with lid loose. When it’s cool close lid and heat seal away filter part of bag. Apply RTV to bag. Contactless “Pour method” from needle.
Have a second small shot glass in the bag as well. Pour LC to that to extract from there. A stack of shot glasses one for each extraction. Cause each time the needle extracts it could be contaminating?
Integrity of jar should be preserved?
reusable only by cutting the bag and adding a new filter to it.
Help me reduce bag waste aspect then we got something.
@@sphericalearther1461 I would say yes, as spores will germinate in nutrient enriched water.