I think this is awesome! I need to test my liquid cultures on agar and this looks like the perfect solution for me. I hate pouring agar in the SAB and then worrying about contamination, and wondering whether it’s from my liquid culture, my agar pour, or the transfer. These have great visibility, as opposed to jars with lids. Does the cotton wick away condensation over time? That would be a bonus. Love that you have no plastic waste and I love reusing discarded glassware, so now to find my alcoholic friends! 😂
Thank you so much I seen a Asian man make a video about this technique and tried to find others trying it as well and came up dry. I then stumbled upon this video, makes me want to try this for sure. It’s a great use of spent glass bottles for sure.
Good thing im the alcoholic relative, now i can just save my own bottles Thanks for this video, ive been looking for an alternative to petri dishes as i can't easily get them locally and i dont like paying absurd prices for jars. Thanks alot !!
Great work. In the interest of thrift I may have to convince a friend or relative to become alcoholic. But seriously, great innovation and nicely presented video practical. I'm just starting off in home mushroom cultivation and am a long way off making my own cultures and spawn but I will try this when I get there. I'll update you at a later date, hopefully. Thanks for this.
Hey Alex , I'm pretty new to all this stuff,but I love learning new things and it's always fun to experiment with different techniques. I'm going to try this , thanks for your video. Take care man, peace ✌️ from Welland Ontario Canada 🇨🇦
Cut a diamond shape maybe will be easier, start at the furthest "point" of the diamond from the mouth of the jar at an outward angle then angle ur blade back inward to complete each side... If that makes sense
When it comes time to inoculate grain and you want to take the whole colony out of the bottle...could be very challenging to do without making a mess... I think this bottle tech applies mainly for observing and sampling mycelium growth when it's difficult to keep out contam (no lam flow). I suppose you could transfer a sample to a small amount of LC prepared in the same way as the agar, then remove mycelium by syringe to inoculate a proper LC jar. For someone with no ability to aquire a laminar flow box I suppose this tech is helpful. 👍
@@DangerRussDayZ6533 Maybe if everyone started with the same resources, skills, knowledge, and teachers. Also, some people like building their own tools and experimenting with different techniques just for the fun of hands on learning. (Edit) LC was the first long term storage tech I tried. It has met my needs with zero hiccups, but my process might be less contam vulnerable than what others do.
Seems like a lot of the scalpel handle is exposed over the agar during transfers, and much of that cannot be flame sterilized. Have you noticed any contamination as a result?
personnaly what I do to prevent that is that I wrap in aluminium foil the tools I'm gonna use (they are dentist tools) and put them in the pressure cooker too for the heat to sterilize them, I only open the foil in my SAB (a glove box actually)
Interesting. Could it be worthwhile to just stick a syringe with liquid culture through the cotton to inoculate the agar in the bottles? Would that not further decrease the chance of contamination aswell as removing the awkward transfer?
I love how unnecessarily complicated this makes things and how it widens the surface area for contamination to be brought in or cause an unrecoverable loss.
Honestly I think that with the opening being narrow and always having the bottle lay flat , in a well prepared GB contamination would be less likely to get onto the agar
@@woutlan I agree, I think the small access to the air above the medium would tend to reduce the contamination. Culture flasks a lot like these are used in professional labs after all
I never had any contams with this technique, it remove the need or pouring the plates after sterilsation wich is a huge bonus if you don't have a workspace that is clean enough or a flowhood
I was wondering if it was possible to warm up the agar without damaging the mycelium and just pour the inoculated bottle into your spawn... Do you guys know the temperature threshold that is bad for mycelium ?
I tried the ketchup cups too, but only the cups themselves are pp5, the lid is other type of plastic so I can’t PC it. I’ve had a high contamination rate and this bottle method seems interesting. I’ll definetly be trying it when I get enough empty bottles to fill up the pC
This is great, thanks for sharing! My only questions, are if agar is blue how can you notice any contamination? An secondly, about how long does it take for your bottles to colonize?
Hi Doug. It's still quite easy to see any contamination. I probably should have shown the bottles held up to the light. The agar is still quite transparent so you can spot different shades of growth. I'd say it normally takes about 2 weeks for a bottle to colonise if it's from a slice of agar from another bottle. Or maybe 3 if it's from a piece of tissue from a mushroom.
This looks insanely hard to sample from, and especially inconvenient because a very specific type of bottle needs to be used. I like pp5 screw top bead containers… they’re cheap and reusable and you don’t have to pour them. After sterilizing
I bought some bottles and became an addiction to alcohol. After that I didn't turned the pressure cooker down and all the agar vaporized... getting stupid from that alc
You could use sterile water spray in in fro. The srynge and use the needle to scrape off the mycelium if waster didn't wash it off then draw it back and you have a culture srynge that can go further
@@alexstacey I did it with oyster on 3 jars. And agar transfer on 4 jars. Agar was growing straight away but the srynge of culture took about 4 days then took over.. I have full colonized grain from the srynge tek. The agar seems to have half stalled
@@Frankiemind recently I found a thing in pharmacy Its sterile stool sample vial with agar in it. Price about EUR 0.5 "tube with transport media" made by equimed company.
I think this is awesome! I need to test my liquid cultures on agar and this looks like the perfect solution for me. I hate pouring agar in the SAB and then worrying about contamination, and wondering whether it’s from my liquid culture, my agar pour, or the transfer. These have great visibility, as opposed to jars with lids. Does the cotton wick away condensation over time? That would be a bonus. Love that you have no plastic waste and I love reusing discarded glassware, so now to find my alcoholic friends! 😂
It supports gas exchange, so yes.
Culture flasks are a revolutionary technique for the at home mycologist. Hopefully one day it'll be brought into the mainstream..
Thank you so much I seen a Asian man make a video about this technique and tried to find others trying it as well and came up dry. I then stumbled upon this video, makes me want to try this for sure. It’s a great use of spent glass bottles for sure.
Good thing im the alcoholic relative, now i can just save my own bottles
Thanks for this video, ive been looking for an alternative to petri dishes as i can't easily get them locally and i dont like paying absurd prices for jars. Thanks alot !!
Great work. In the interest of thrift I may have to convince a friend or relative to become alcoholic. But seriously, great innovation and nicely presented video practical. I'm just starting off in home mushroom cultivation and am a long way off making my own cultures and spawn but I will try this when I get there. I'll update you at a later date, hopefully. Thanks for this.
Is there a reason for using 200ml and not 100ml??
BTW: I love this, thak you for the tek!
How innovative. Thanks for bringing this to our attention. I do a similar thing but with 1/2 pint mason jars. Good job Alex.
This is so freaking cool!!!
danke..
Hey Alex , I'm pretty new to all this stuff,but I love learning new things and it's always fun to experiment with different techniques. I'm going to try this , thanks for your video. Take care man, peace ✌️ from Welland Ontario Canada 🇨🇦
I use this technique too. The best
Cut a diamond shape maybe will be easier, start at the furthest "point" of the diamond from the mouth of the jar at an outward angle then angle ur blade back inward to complete each side... If that makes sense
I'm doing this now and i have to say its brilliant bro
When it comes time to inoculate grain and you want to take the whole colony out of the bottle...could be very challenging to do without making a mess...
I think this bottle tech applies mainly for observing and sampling mycelium growth when it's difficult to keep out contam (no lam flow). I suppose you could transfer a sample to a small amount of LC prepared in the same way as the agar, then remove mycelium by syringe to inoculate a proper LC jar.
For someone with no ability to aquire a laminar flow box I suppose this tech is helpful. 👍
If you're able to afford a laminar flow, you probably should know better than to be working with LC in the first place ;)
@@DangerRussDayZ6533 Maybe if everyone started with the same resources, skills, knowledge, and teachers. Also, some people like building their own tools and experimenting with different techniques just for the fun of hands on learning.
(Edit) LC was the first long term storage tech I tried. It has met my needs with zero hiccups, but my process might be less contam vulnerable than what others do.
Oh My God. that's genius thank you so much saved my life and gave a bunch of money to my corner shop lmao those poliakov bottles man...
I love this! Thank you!
Stumbled on this today,gold!!
Seems like a lot of the scalpel handle is exposed over the agar during transfers, and much of that cannot be flame sterilized. Have you noticed any contamination as a result?
personnaly what I do to prevent that is that I wrap in aluminium foil the tools I'm gonna use (they are dentist tools) and put them in the pressure cooker too for the heat to sterilize them, I only open the foil in my SAB (a glove box actually)
Love how well you explain this 🌟🌟🌟
Interesting. Could it be worthwhile to just stick a syringe with liquid culture through the cotton to inoculate the agar in the bottles?
Would that not further decrease the chance of contamination aswell as removing the awkward transfer?
This is badass
Very cool thanks brother.. I was just about to spend coin on Pyrex petri dishes think I'm going to try the bottles .. peace
I love how unnecessarily complicated this makes things and how it widens the surface area for contamination to be brought in or cause an unrecoverable loss.
Honestly I think that with the opening being narrow and always having the bottle lay flat , in a well prepared GB contamination would be less likely to get onto the agar
@@woutlan I agree, I think the small access to the air above the medium would tend to reduce the contamination. Culture flasks a lot like these are used in professional labs after all
Bareknuckle you miss the point of this method and sell assumptions as facts.
Completely unnecessary comment
I never had any contams with this technique, it remove the need or pouring the plates after sterilsation wich is a huge bonus if you don't have a workspace that is clean enough or a flowhood
I was wondering if it was possible to warm up the agar without damaging the mycelium and just pour the inoculated bottle into your spawn...
Do you guys know the temperature threshold that is bad for mycelium ?
No u cant itll for sure damage the mycelium
this is awsome mate, thanks, i already brought up some ketchup cups for alternatives
I tried the ketchup cups too, but only the cups themselves are pp5, the lid is other type of plastic so I can’t PC it. I’ve had a high contamination rate and this bottle method seems interesting. I’ll definetly be trying it when I get enough empty bottles to fill up the pC
Thanks for sharing.
How long do you wait to let the pressure cooker depressurize?
Pretty good technique!
This is great, thanks for sharing! My only questions, are if agar is blue how can you notice any contamination?
An secondly, about how long does it take for your bottles to colonize?
Hi Doug. It's still quite easy to see any contamination. I probably should have shown the bottles held up to the light. The agar is still quite transparent so you can spot different shades of growth. I'd say it normally takes about 2 weeks for a bottle to colonise if it's from a slice of agar from another bottle. Or maybe 3 if it's from a piece of tissue from a mushroom.
This is so cool
I bc wanna see u struggle to get that out to inoculate grain , u got a video on that??
Clever
You got a link for those bottles
This looks insanely hard to sample from, and especially inconvenient because a very specific type of bottle needs to be used. I like pp5 screw top bead containers… they’re cheap and reusable and you don’t have to pour them. After sterilizing
Does this method help get rid of condensation with those cotton balls?
Try a #7 handle with a # 11 blade 😊
Check out the glass ware from the gu yogurts your love them
I bought some bottles and became an addiction to alcohol. After that I didn't turned the pressure cooker down and all the agar vaporized... getting stupid from that alc
Probably don't hear about it much in Europe because it's odd and is less efficient this way.
Shout out the the "alcoholic relative" 😅
you can remelt agar, and pour it into a trash....for cleaning...no need to use that long stick to get pieces out..
I hope I’m not to late to the party, how do you go about inoculating grains? Remove tiny pieces?
One question, after my plates are fully colonised should i store them in the fridge or keep them at room temp?
how do you get them out for next transfer?
I just do the same method with a skalpel
You could use sterile water spray in in fro. The srynge and use the needle to scrape off the mycelium if waster didn't wash it off then draw it back and you have a culture srynge that can go further
@@johnpwright7832 that's a really interesting idea! I'm going to try that.
@@alexstacey I did it with oyster on 3 jars. And agar transfer on 4 jars. Agar was growing straight away but the srynge of culture took about 4 days then took over.. I have full colonized grain from the srynge tek. The agar seems to have half stalled
I’d have such a hard time doing that
you sound like david walliams but cool vid
Why would you do this when jars exist? Just 4oz jam jars man...
Less potential air movement in a narrow bottle than in a wider jar
Y’all will do anything but buy regular Agar plates
Baby food jars bro
Their jars lid wont close after putting it under pressure cooker... how do you work with lids on your bay food jars?
@@welldun9769 go on Amazon, look up the Amersumer plastic bead containers. These are the best for agar.
Dishes are cheap no need to make life harder
Thats exactly what i thought. Please just work cleaner...
@@Frankiemind recently I found a thing in pharmacy Its sterile stool sample vial with agar in it. Price about EUR 0.5 "tube with transport media" made by equimed company.
@@Krzemieniewski1 thats very interesting. 🧐
Jesus just use 4 oz jars