HOW TO MAKE LIQUID CULTURE FOR MUSHROOM GROWING, Non-Vented Lid Jars from Start to Finish

[@RenegadeMushrooms]

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: kzbin.info/www/bejne/hKeqiIOVn6uNo9E

[@ChristophHalverson-iw8gk]

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!

[@ChristophHalverson-iw8gk]

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?

[@RenegadeMushrooms]

@@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.

[@benchadwick1754]

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!

[@malourdessun]

Thank you very much for sharing your expertise. Much appreciated ❤

[@samgodfrey8141]

Thanks for the fast replies, your 20 yrs experience is a great asset . I appreciate it.

[@rickfoss1136]

Best mushroom channel on KZbin. 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?

[@RenegadeMushrooms]

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.

[@rickfoss1136]

@@RenegadeMushrooms thanks for the reply! Keep up the great work and thanks for sharing all your knowledge!

[@vk081064]

This is by far the best video on liquid culture. Thanks for your efforts to educate beginners like me

[@RenegadeMushrooms]

Appreciate you watching and commenting. Let me know if you have any questions.

[@HeyRaeSunnyDay]

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.

[@RenegadeMushrooms]

Glad you enjoyed the video 👍 I'm always here if you have questions. Thanks for watching and subbing.

[@Unclemushlovin]

Cheers new to growing mushrooms looking for good info that you have provided thanks mush lovin

[@smartjared7203]

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[@smartjared7203]

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[@francisr6804]

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?

[@RenegadeMushrooms]

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 👍

[@francisr6804]

@@RenegadeMushrooms Awesome, let me know if it works!

[@bestheirs8999]

@@RenegadeMushrooms Can one use plastic jars instead of glass jar?

[@RenegadeMushrooms]

@@bestheirs8999 Plastic jars are ok as long as you use thick PP5 plastic, but glass is superior in my opinion.

[@davidjackson552]

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.

[@RenegadeMushrooms]

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.

[@glennplatvoet7111]

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 .

[@RenegadeMushrooms]

Awesome, having a flowhood is a game changer.

[@aidennymes6335]

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!

[@RenegadeMushrooms]

Good idea, thanks for watching

[@addammadd]

(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.

[@RenegadeMushrooms]

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 👍

[@loricox3833]

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.

[@kevinabood3598]

thank you for this i was searching for how to make a diy injection port

[@johnhall9378]

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)

[@RenegadeMushrooms]

I find both are useful, so it's good to know how to make vented and non-vented LC.

[@bmartin2304]

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!

[@mattnsim]

Never new about master jars! Thanks mate

[@FlomatonFamous]

Good stuff, good info in your video's

[@glennplatvoet7111]

That's slick thank you

[@SirBoden]

Minor point: Heat sterilizes, alcohol sanitizes. Wiping a heat sterilized needle with alcohol downgrades it to sanitized. This is good enough in this situation.

[@jasonedwards2622]

Valid point. I like to pressure cook a jar of syringes with my liquid culture

[@C0Y0TE5]

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...

[@Alibali50]

Do you have a video that shows how to withdrwal from the jar in the best way? 😮

[@RenegadeMushrooms]

Just tilt them upside down 45 degrees and withdraw the plunger on the syringe. You can also just lay the jar on it's side if it's pretty full.

[@Junzig]

Great video. Thank you

[@noelmansfield8505]

Thank you! This was awesome!

[@seanvan6377]

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.

[@RenegadeMushrooms]

Cool trick 👍 This alcohol soaked polyfil deal is pretty quick and effective as well.

[@TheDMTLover]

Good tricks. Thx. 🙂

[@J155P]

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.

[@RenegadeMushrooms]

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.

[@dudetapedtoafridge3073]

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

[@dudetapedtoafridge3073]

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!

[@RenegadeMushrooms]

Good tip, thank you 👍

[@MiigsMusic]

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.

[@RenegadeMushrooms]

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.

[@jbb3141]

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.

[@RenegadeMushrooms]

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.

[@Rhizomorphius-vz4bj]

I noticed you equalized the pressure inside the jar then injected 10 CCs of LC going from negative pressure to a slight positive pressure inside the jar, meaning you put 10 CCs more air in the jar than you actually needed. If you need to introduce an estimated 30-40 CCs of air, why not start with injecting your 10 CCs of LC first, thus reducing the amount of air needed to equalize pressure by 10 CCs, and reducing your chances of contamination by doing so. It just makes more sense to put the LC in first then relieve the remaining vacuum.

[@RenegadeMushrooms]

Never measured it and it varies by jar but I would guess it's more like 100 - 200 ccs to initially equalize a pint LC jar. I usually only add 3 or 4 ccs from the syringe. If you injected a jar in a state of full vacuum it would rip a 10 cc syringe plunger down instantly. In theory I get what you're saying, but in practice once you do the initial equalization everything else works itself out in the wash.

[@malekith8215]

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.

[@RenegadeMushrooms]

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.

[@malekith8215]

@@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. 😉

[@raulduke1528]

Very nice

[@chillville5571]

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?

[@RenegadeMushrooms]

Two injection ports isn't really necessary. I always look at it like any additional holes in your lids are potential contamination points.

[@angel2jimenez219]

@@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

[@RenegadeMushrooms]

@@angel2jimenez219 Not necessary.

[@glennplatvoet7111]

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

[@RenegadeMushrooms]

There are lots of designs out there. I prefer a centrifugal fan setup myself. I explain how I built my hood in this video: kzbin.info/www/bejne/d3acpYNsZq-ahpo

[@jaydenallanach969]

Do you think the pickle jar lid and a hole with medical tape would work?

[@RenegadeMushrooms]

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.

[@samgodfrey8141]

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.

[@RenegadeMushrooms]

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 👍

[@ginalogist]

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

[@RenegadeMushrooms]

I'm actually using a 16 gauge BD needle that I got for free with a LC syringe.

[@FelixNothus]

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?

[@RenegadeMushrooms]

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.

[@FelixNothus]

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.

[@MrLuong]

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?

[@RenegadeMushrooms]

Yes, it is to clean any potential contaminants out of the syringe and needle.

[@AM2PMReviews]

How do you pressure sterilize with an instant pot?

[@RenegadeMushrooms]

I've never used an Instant Pot so I don't know. I'm sure you can find some vids on YT.

[@AM2PMReviews]

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

[@RenegadeMushrooms]

@@AM2PMReviews Lions Mane vid: kzbin.info/www/bejne/hF60oZVpgJ14gpI 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.

[@mjarami3]

Would you use this method for a bigger jar? Or would the larger jars require more fresh air?

[@RenegadeMushrooms]

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.

[@byankashomestead8038]

Great video! I was wondering if you had one on agar? I'm having a lot of contamination and wonder what you do. ☺️

[@RenegadeMushrooms]

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. kzbin.info/www/bejne/fWO2mpykqKqWeas

[@byankashomestead8038]

@@RenegadeMushrooms Great thanks!

[@JohnLeggio]

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

[@RenegadeMushrooms]

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.

[@JohnLeggio]

​​@@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

[@jmichel70]

Thank you

[@JasonAlexzander1q47]

Will that work with a syringe full of spores? Inoculate a jar with a syringe full of spores?

[@RenegadeMushrooms]

Yes it works well with spores too as long as the spore syringe is clean and sterile.

[@lizzp9830]

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?

[@RenegadeMushrooms]

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.

[@bqoutdoors2334]

When you pressure cook the jars do you tighten the lids? Also do you turn the lids upside down? Thanks I subbed

[@RenegadeMushrooms]

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.

[@kumhoshin8736]

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?

[@RenegadeMushrooms]

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.

[@kartalbaba981]

Thank you!

[@superagentHH]

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 🙏

[@RenegadeMushrooms]

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.

[@AlexanderSchwarz-ck3cx]

When you put the squeezing thing out of the syringe, isnt contaminated air going inside?

[@RenegadeMushrooms]

@@AlexanderSchwarz-ck3cx No, any air that flows through the syringe into the jar is filtered by the isopropyl soaked polyfil inside the syringe.

[@AlexanderSchwarz-ck3cx]

@@RenegadeMushrooms ahh and its automatically flowing, because there is a vacuum inside the jars. Thank you for the quick Response, I definitely subscribe!

[@loricox3833]

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

[@RenegadeMushrooms]

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.

[@bg31337]

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

[@RenegadeMushrooms]

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.

[@johnhall9378]

Could you just use a vented lid, then after the jars cooled and pressure equilibrated, seal the filter disk with tape or RTV ?

[@RenegadeMushrooms]

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: kzbin.info/www/bejne/hKeqiIOVn6uNo9E

[@lycanzhp1141]

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

[@RenegadeMushrooms]

It's more like 200+ ccs to equalize.

[@lycanzhp1141]

@@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 :)

[@RenegadeMushrooms]

@@lycanzhp1141 Awesome, glad it worked for you 👍

[@serenity6988]

Great video man!! ?? Where do I find the 12 ga needle? I have looked every where. Can you send link please.

[@RenegadeMushrooms]

I've had trouble finding them myself lately, but I know Shroom Supply has 14 gauge needles and they work well.

[@myotheraccount5947]

I see a lot of videos where guys are using magnetic stirrers to help the mycelium grown evenly throughout the liquid. Not necessary?

[@RenegadeMushrooms]

Not necessary, but a very nice tool. I've never bought one because I've never really found I needed it, but they are cool.

[@myotheraccount5947]

@@RenegadeMushrooms Thank you!

[@DrGaryGreen]

Closed, non-vented lid. PC'ed and the injection port doesn't blow and the glass jar doesn't explode? Ok..... wow. I've been doing the quarter to half turn to loosen the lid during PC'ing to prevent jar explosions. And I don't have to? Bye bye to another possible source of contamination. Thank you sir.

[@RenegadeMushrooms]

I have to be careful because the PC police love to verbally abuse me, but I've never had an LC jar break, crack, explode, etc. But I only PC LC jars for 25 minutes at 15 psi. I just snug the lids down and they actually seem to loosen a bit on their own during cooking. I always resnug them down as soon as I pull them out.

[@DrGaryGreen]

@@RenegadeMushrooms Thank you for your reply and words of experience. I'm going to follow your lead and if anything cracks I'll respectfully let you know.

[@robertmenucci2668]

Can you reuse all syringes, from the same strain, to transfer LC? Like taking LC out to Vax substrate?

[@RenegadeMushrooms]

I use a syringe once, pulling in LC and then dispensing it into jars. IMO pulling more LC back in risks introducing contamination without resterilizing the syringe. Not saying you couldn't get away with it but I don't do it. Syringes can be washed, wrapped in foil or placed in a jar, and re-sterilized in the PC though. I also use 20cc syringes when doing transfers so I get a little more volume. Syringes with LC left in them can be stored for a couple months in a ziploc bag.

[@robertmenucci2668]

Thanks for the reply. I'm a big fan of the show. Being new at this, I have to "pick my battles" to save money, and I'm not willing to risk an entire batch in order to save a few bucks

[@mtrobenazz]

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?

[@RenegadeMushrooms]

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.

[@RudolfAtterdag]

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.

[@RenegadeMushrooms]

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: kzbin.info/www/bejne/aGnYhpyZfJuAaMk

[@DEeMONsworld]

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?

[@RenegadeMushrooms]

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.

[@DEeMONsworld]

@@RenegadeMushrooms yes every weak link... I'm small time just starting out, looking for ways to prolong liquid culture if I get that far

[@calichojack991]

Do you need a torch or could you do this with a lighter?

[@RenegadeMushrooms]

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.

[@danherring5676]

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.

[@RenegadeMushrooms]

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.

[@mechamotollc]

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?

[@RenegadeMushrooms]

Yes that would work too. I've done that before with the foil wrap and it worked well.

[@irate74]

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?

[@RenegadeMushrooms]

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 👍

[@mse1333]

Hey Renegade…..can you use spores to colonize your liquid culture? Love your vids!

[@RenegadeMushrooms]

You can as long as the spore syringe is sterile, it works great. Thanks for watchin 👍

[@mse1333]

@@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.

[@mattnstewart262]

Could one stuff a synthetic filler disk in the syringe instead?

[@RenegadeMushrooms]

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.

[@Magicsam87]

Can you not just suck the air back in infront of the flow hood to get sterile air?

[@RenegadeMushrooms]

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 😁

[@crypto.entrepreneurs]

Thanks for sharing your knowledge. I was wondering if I can replace polyfil with cotton?

[@RenegadeMushrooms]

Cotton, being a natural fiber, absorbs water and is more prone to contamination. Polyfil is cheap and definitely the way to go.

[@crypto.entrepreneurs]

​@@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

[@RenegadeMushrooms]

@@crypto.entrepreneurs You can also use 2 layers of micropore or cloth medical tape if you can find that.

[@crypto.entrepreneurs]

@@RenegadeMushrooms Yes that I have. I'll try it that way too. Thanks

[@zumbapatty4988]

@@crypto.entrepreneurs I found it in the sewing section of Walmart - get the fiber fill, not the sheets

[@DukeTurnip]

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?

[@RenegadeMushrooms]

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.

[@calichojack991]

Can I buy syringes in local stores or do I need to order them online?

[@RenegadeMushrooms]

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

[@nobletravistn]

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.

[@RenegadeMushrooms]

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.

[@timm8311]

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.

[@RenegadeMushrooms]

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.

[@kimelize]

How long can you keep a master jar going?

[@RenegadeMushrooms]

1 year or a bit more under refrigeration.

[@wickedjack1222]

Wouldn't just squirting (the jar actually sucks) the LC syringe equalize the pressure inside the jar ?

[@RenegadeMushrooms]

Not even close, you could dump a full 20cc syringe in and still have vacuum.

[@cliffmello]

Hey boss, were can I get empty syringes? You order online? Thank man. Keep up the great vids

[@RenegadeMushrooms]

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.

[@Beyondbuttonmushrooms1738]

Where did you manage to find a 12 Gauge syringe tip? Im not having a lot of luck.

[@RenegadeMushrooms]

They're hard to find nowadays. Shroom Supply has 14 gauge needles the work great though.

[@Beyondbuttonmushrooms1738]

@@RenegadeMushrooms I wonder why this is the case. Is preferred amongst drug abusers? 🤔

[@trishdeon4313]

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 :)

[@RenegadeMushrooms]

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.

[@ymm424]

what size are those black self-healing plugs please? I just searched for them and found they come in two sizes.

[@RenegadeMushrooms]

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%.

[@Jason-oy6vp]

@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.

[@RenegadeMushrooms]

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.

[@Jason-oy6vp]

@@RenegadeMushrooms Thank you! I will.

[@rfyock009]

Ty so much! What micron is micropose? The reason for the poly fill syringe?

[@RenegadeMushrooms]

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.

[@saintlucidVR]

thanks man

[@philq01]

I noticed you do not use magnetic stirers. Is that not necessary

[@RenegadeMushrooms]

They are great, but not necessary. I just shake/swirl my jars by hand and it's always turned out fine. I would say go for it though if you have the money.

[@stayanddrown]

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!

[@RenegadeMushrooms]

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 👍

[@stephenwalther9702]

​@@RenegadeMushroomsaway another question, please: why couldn't you tape over a vented lid filter with regular tape, to prevent further air exchange?

[@RenegadeMushrooms]

@@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.

[@rickyonoroto2094]

Can spore syringes be used with method?

[@RenegadeMushrooms]

Yes, as long as the syringe is sterile.

[@blenderbenderguy]

I have the same torch..... I keep knockin it over.... it's time for a smallish alcohol lamp.

[@RenegadeMushrooms]

Dang lol, I've never knocked mine over. But you do definitely have to go with the fatty propane bottle if you're going to rock that torch for the stability factor.

[@faqu4862]

Would a stopcock work for equalizing the pressure?

[@RenegadeMushrooms]

Yes, if it was mounted to the lid, and you used some type of sterile filter media.

[@bigdogdazza]

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?

[@RenegadeMushrooms]

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.

[@bigdogdazza]

@@RenegadeMushrooms awesome thank you!

[@Fall3nL3g3nd]

Do you think if i make it with Tyndalization instead of pressure cooker will it be fine?

[@RenegadeMushrooms]

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 👍

[@samgodfrey8141]

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.

[@RenegadeMushrooms]

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.

[@samgodfrey8141]

@@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.

[@RenegadeMushrooms]

@@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.

[@goodelleric]

@@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…

[@RenegadeMushrooms]

@@goodelleric You might be better off just having a filter disc in your lid until you have a flowhood setup.

[@mattemmerson5877]

Do you reuse the foil lids? I never thought about that.

[@RenegadeMushrooms]

Yes, I re-use them many times, until they get brittle.

[@archieh4058]

Are the RTV lids dishwasher safe?

[@RenegadeMushrooms]

Yes I would think so, but I always hand wash them, only take a second.

[@J155P]

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

[@RenegadeMushrooms]

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.

[@DukeTurnip]

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.

[@RenegadeMushrooms]

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.

[@mse1333]

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!

[@RenegadeMushrooms]

@@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.

[@daveeatmon4718]

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.

[@mse1333]

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?

[@RenegadeMushrooms]

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.

[@RobBanks81]

If the jars were filled almost completely, would there be enough oxygen for a year's storage?

[@RenegadeMushrooms]

@@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.

[@diverwa]

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?

[@RenegadeMushrooms]

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.

[@diverwa]

@@RenegadeMushrooms Thanks for the follow-up. One more question, have you ever tried cooking your grain in agar befor inoc to speed up spawn?

[@RenegadeMushrooms]

@@diverwa No I haven't, but let me know how it works if you try it.

[@haroldjoseph3596]

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.

[@RenegadeMushrooms]

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.

[@professored7169]

If you have have a pc then do no pour agar, its what I do because I use a SAB

[@toddwmac]

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. :)

[@ChristophHalverson-iw8gk]

I’ve had a bit of bad luck with my LC coming out of the PC looking like a dark burnt mess, any tips?

[@RenegadeMushrooms]

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.

[@ChristophHalverson-iw8gk]

@@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

[@RenegadeMushrooms]

@@ChristophHalverson-iw8gk What recipe are you using, mine?

[@Akasha_Silver]

Hey,how much water & Karo syrup would I use to make only 2 pint jars? Thanks

[@RenegadeMushrooms]

You want a 4% solution so just take your total distilled water volume and multiply it by .04, that will be your amount of karo. Not perfect but close enough 😁 I usually put 300 ml. of liquid in my pint jars so 300x.04=12. So add 12 ml. of karo to each jar if you go with 300 ml. You can go a hair over that without hurting anything.

[@Akasha_Silver]

@@RenegadeMushrooms thanks

[@davidblack5551]

The. MASTER 👍

[@jeremyprince4945]

Do u have to use the soy peptone?

[@RenegadeMushrooms]

No, it's a nice supplement. But just karo will work.