I expanded on this idea by making vents and injection ports in the lids. First, if you decide to add the agar to the jars before sterilizing make sure you cool it very slowly. My way: 1. Same jars but I drill two holes in the lids: one 1/4 inch and on 1/16 inch. 2. Grab a small wad of poly fill and fold it in half. Then use a 1/32 drill bit or similar sized pin to push the poly fill through the 1/4 inch hole. Grasp the tiny bit of poly fill on the other side of the hole with your fingers and gently and firmly keep pulling the poly fill through the hole by re grabbing it close to the hole. It will become thicker and thicker until it is difficult to pull through. I use a circular motion to get the poly fill good and tight. Then trim away the excess poly fill from the inside of the lid so that it only has about 1/4 inch showing. I like to leave about a 3/4 inch ball of the poly fill on the outside of the lid. 3. Get some blue permatex RTV gasket and squeeze a small 1/2 inch ball into the 1/4 inch hole. Dab your finger in some oil and gently push the ball back and forth through the hole so the sealer sandwiches the lid on each side of the hole. Make sure it is about 1/8 inch thick. Let the lid/s dry for 24 hours. 4. Add the correct amount of agar solution to each jar and screw the lids on and tighten lightly. 5. Put a piece of tinfoil over each jar. 6. Sterilized the jars in a PC at 15 PSI for 15 minutes. Be sure to add enough water to submerge the jars about 3/4 inch. Heat up and cool down slowly to avoid rapid boiling the PC and boiling over the jars. 7. You now have vented petri dishes that can be easily inoculated via syringe through the permatex self healing injection ports. If done carefully you should not need a glove box to inoculate.

@living4adrenaline

5 жыл бұрын

CAVEAT: Spore syringes are mostly water which could introduce contaminants. Spore print to agar via inoculation loop, sterilize between jars

@lilkaps3344

5 жыл бұрын

@@living4adrenaline I've drilled 1/4 hole in my elite canning jars and instead of polyfil I'm using SFD cut into circles with a rtv ring. Will this work?

@lilkaps3344

5 жыл бұрын

@@living4adrenaline bump

@mauialejandre6370

4 жыл бұрын

Sir what is the content of your agar solution

@Sgtspork

4 жыл бұрын

micropore medical tape works just as well as polyfill and is a lot more easy to handle overall and no worries about grain or substrate getting caught up in it on the underside of the lid.. also, straight silicone sealant from a tube self seals as an injection port just fine. use the mason jar ring to hold down the tin foil... just my experience.

Why not mix the agar, pour into small jars and THEN sterilize? That's what I do and I never have contams. Just my 2 cents

@shawnewing3971

7 жыл бұрын

my 2 cents... no significant problem sterilizing in "jars" (no pour). You do have to contend with boil overs, or water getting into jars. This method basically mimics the standard method of pouring into petri dishes. In this case substituting jars. Petris make it easier to identify contam's, but they are not pressure cooker friendly (when loaded with agar - dry is ok tho).

@tonysimonsen3228

6 жыл бұрын

HDRWIT im certainly not going to try arguing his method is wrong, all i can say is i make mine up in a regular saucepan. I get it to a boil, leave it (covered) to cool down, then pour into my petri dishes. A lot less effort and it hasnt failed me yet

@kushdog8

6 жыл бұрын

With the materials given “no pour” is a much better option. You get better results with less work using the materials in the video. Hell, you wouldn’t need to open any jar or bust out the sab/glove box until your ready to inoculate.

@andersonwade9963

5 жыл бұрын

Hi can you tell me how long should the agar be sterilize in the pressure cooker?

@williamcozart9166

5 жыл бұрын

It's usually for 15-30 minutes at 15 psi. Like the comments above said, using glass jars like in the vid instead of plastic pitris, you can mix the agar and malt and pour into jars before pressure cooking, then you can skip the sterile "pour" step.

Would it save time to first measure the malt extract into the primary mixing jar first, then add the measured water and stir. ...then pour in the agar? The extra step is adding work and dishes. This way you never need to clean malt extract off anything.

@lisak7545

3 жыл бұрын

It would also make sure that 1 of the 10 grams is not left behind on the vessel.

really good video, thanks

Good job! Thanks :)

very nice concept with your own lab

Greetings, Well made video, thank you. Question: Prior to the sterilization process can one go ahead and separate the agar into the canning jars and then proceed with sterilization as if one was canning? The jars would be sterilized and sealed preventing contamination and preventing the need for the glove box and transfer operation. Just a thought.

@ignaciocabero6062

4 жыл бұрын

If you're talking about using the boiling sealing method for "sterilizing" cans then it wont actually sterilize. Bacterial spores or other contaminants can still survive the boiling sealing method so using a pressure cooker (ideally an autoclave) is essential

@cindyajohnson267

3 жыл бұрын

@@ignaciocabero6062 No, not necessarily. It would be pressure canning. The jars would be kept sterile until used. So you wouldn't flip the lids, you'd seal them. Just like you were pressure canning a low acid jelly or jam.

Thanks for the video! Can I use molasses instead of malt extract?

You need to put a metal plate or towel on the bottom of the pan before putting the jars in to prevent the heat from the stove breaking the jars. They have special plates for the bottom of pressure canners. It's very important to use them, or you can have glass everywhere!

Mine still boiled over and 3/4 of the liquid was gone when I opened it ...

Could you place those petri dishes back in the pressure cooker for a second time before placing them in the refrigerator?

excellent, thankyou xxx

nice vid.. tnx

Freaking awesome 👌 👏

Thanks brother. How long it can be stored usually? And what is the benefit of using mason jar rather than petridishes?

@ThePinkBinks

2 жыл бұрын

The jars are reusable. I would do the No Pour method with the jars though since they need to be cooked either way and there’s less chance of contamination that way. There’s also less chance of any jars cracking because they’ll cool down slower. Sauce pots made from PP plastic work as cheap petri dishes just fine and they survive the pressure cooker.

What is the difference when dextrose is used? I see people using that. Is it just a different form of sugar?

I seen online this recipe calling for dextrose? Wouldn't powdered dextrose and nutritional yeast boost growth rates?

I want to know how to grow hair using a Petri dish. Is this possible? I want to try adding different ingredients with a hair strand in each Petri dish. Can you please give some suggestions?

I would assume any type of sugar could replace the malt extract at the right ratio?

Would it be wrong to sterilise agar (pre-poured) in the jars?

Better to weight the malt extract in your mixing jar, so you don't have to try and pour and scrape into it,

My pressure cooker is standard and i dont know how to use it, HELP

"Reynolds Wrap" .... "Reynolds" Hamilton: TRIGGERED

Does the malt extract have to be "Light"? AWESOME vid thanks!

@Sgtspork

4 жыл бұрын

there's premix stuff that works great and is cheap.. check amazon for MEA agar...

Hallo can you use this mix you make for orchid en plant to grow inside

does this recipe work well for growing bacteria

Can malt extract be replaced with table sugar?

yes ur right! he could have just put the liquid in the small jars and the steam will sterilize the jar, the liquid and the lid. the lid has glue on the gasket if it a new one. 15psi = 250 degrees F. he could have used honey instead of malt also!

@daamu48

5 жыл бұрын

Can we heat honey ?

@taylorhitchcock5585

5 жыл бұрын

joe smith could one substitute the malt with kyro ?

@andersonwade9963

5 жыл бұрын

Hi can you tell me how long should the agar be sterilize in the pressure cooker?

@taylorhitchcock5585

5 жыл бұрын

@@andersonwade9963 20 minutes at 15psi

@living4adrenaline

5 жыл бұрын

@@taylorhitchcock5585 30 - 40 @ 15 PSI

Also Doesn’t mycelium need to breath? There was a seal and no sterile gas exchange using micropore or polyfill stuffing

Your Agar doesn’t go bad for not leaving it upside Down?

Did you have to make holes in the tops of the jars?

What is the point of pouring the liquid inside the glove box if you're going to open the jar outside of the box. Wouldn't that defeat the purpose of sterilizing the jars,causing possible contamination?

Can I use anything else instead of that malt?

where can i get the jars withe the "flipable lid"? i cant find them. Is there any special name for this?

@LiborTinka

6 жыл бұрын

search for "wide mouth mason jars" - they are made by Kerr and Ball ... but IMO the flippable lid is not needed, you flip it only to prevent rubber sticking to the glass. I am using regular screw lid just fine. You can add Parafilm or even duct tape later if concerned about the seal.

i followed this tek and its been a week since inoculation and there is no mycelium, are u sure this tek works or do you think i might have done something wrong?

@AlAllerton

5 жыл бұрын

Comment Crafter, (for others asking the same question), spores can take weeks to show growth. Though 10 days is about average. How did yours turn out?

Could you use honey

Can I just put the agar plate/jar inside boiling water if I don't have a pressure cooker?

@fresnomycologysociety5004

2 жыл бұрын

Boiling water alone won't kill everything. You need the pressure and the higher temp you get with it

Can i make this for plant

How to get this jars ?

You don't need malt extract, you don't need distilled water, you need deionized water! It's different. Btw, how long did you take to completely poor the malt extract from the cup into the jar ?!

Can anybody tell me wether they have had improved results introducing a gas exchange hole with a layer or two of micropore tape? I’m going to be making my own agar and can’t decipher from the internet wether this method should have gas exchange. I’ve used Petri dishes and used them in parafilm. But I think the paradigm allows for SOME GE. Any help, helps!

@trevorswayhome

3 жыл бұрын

Parafilm*

@fresnomycologysociety5004

3 жыл бұрын

Yeah I usually drill 1/2 inch hole in the lids and seal a circle of tyvek or filter paper with some hi temp silicone. Trying the same thing in place of poly-fil on a monotub as well.

its proven that attaching gloves to your SAB is more likely to attract contams.

@TURK_182

4 жыл бұрын

Correct, a glove box moves air more

@tajdavis915

3 жыл бұрын

@Thought Frankly I think because when you move the gloves around it creates a 'billowing' effect and sucks air in and out

@funny11744

3 жыл бұрын

@Thought Frankly It îs about pressure and vacuum- but it îs not the case If the dimensions of gloves remain constant.

@morganconklin8849

3 жыл бұрын

I got an sab coming with gloves already attached. It's got great reviews I hope i dont have trouble with it.

@Lurkspur

3 жыл бұрын

@@morganconklin8849 just cut em off! Trust me, gloves love to attract contamination

Why not pour in little jar before boiling?

What is use cultue??

Obviously agar is needed. Not many of us keep light malt extract in the pantry, are there any alternatives? Honey? Awesome video

@pequodexpress

5 жыл бұрын

I am looking for an all-in-one dry agar mix. Not sure if I will know what it is if I find it.

@living4adrenaline

5 жыл бұрын

@@pequodexpress See this video: kzread.info/dash/bejne/gY1206StZ7LLgtY.html 500ml water to 20g agar should be enough

@pequodexpress

5 жыл бұрын

No need to add salt, Triptose, or anything else? I am doing this only to play around with some bacteria experiments with my seven-year-old son

@living4adrenaline

5 жыл бұрын

@@pequodexpress Actually I believe RR, guy in the video linked was using agar mix, probably with malt extract as this guy is. Honey should work in theory

@living4adrenaline

5 жыл бұрын

@@pequodexpress Cool! I recall an experiment in us where we wiped our fingers across wet Petri, left em for a bit and they were loaded with bacteria

Can you use Gelatin

Please mention what dark liquid îs in the square recipient at 0:30 minute.Thanks

@fresnomycologysociety5004

3 жыл бұрын

Light malt extract

@funny11744

3 жыл бұрын

@@fresnomycologysociety5004 I found malt extract , but how I get LIGHT malt extract ? Any idea ?

@fresnomycologysociety5004

3 жыл бұрын

@@funny11744 www.midwestsupplies.com/products/organic-light-lme-maillard-malts?variant=30973169795146

Defined media? Or complex?

can I use this setup for Ecoli growth?

@fresnomycologysociety5004

3 жыл бұрын

Totally, just might want to use an LB agar mix instead of malt extract.

Great video and great channel, thank you! The music covers your voice to the level of making it occasionally difficult for me to hear what you say, could you please change the balance in favour of your voice?

@fresnomycologysociety5004

3 жыл бұрын

Glad you enjoyed it! These videos were a lot of fun and a huge learning experience for producing quality stuff. Our future videos will try to dial in the mix better!

Can i just honey in place of malt?

@ignaciocabero6062

4 жыл бұрын

You can use potatos and honey, but I don't think honey itself will work. Look up how to make potato dextrose agar or something similar

Can I do this with spore syringe?

@hydrolink512

5 жыл бұрын

Of course you can, but you are creating a potential source of contamination, the agar is already hydrated and provides the necessary moisture for the mycelium. Scraping the spores on the agar would suffice.

thank you...the music is very loud, in some parts the voice is not heard well.

@fresnomycologysociety5004

Жыл бұрын

For sure. This was one of the first videos we made, we were still developing our production skills haha thanks for the feedback!

This jar much better then plastic

why dont you sterilize directly in the jars? you would skip the pouring and you could even use modified lids with injection port.

@fresnomycologysociety5004

Жыл бұрын

You got a good brain, friend. You're right! You could defs do that, this was one of the earliest videos I had made, i was still learning a lot.

I gotcha. I should just buy them. He said a fews things and pulled out 100 things. I-

Why not just put the jar on the scale and zero it out after each addition?

@bartosznaporowski6383

5 жыл бұрын

if you add too much, you can remove some.

How many liters of autoclave?

@Sgtspork

4 жыл бұрын

no more than halfway up the side of the bottom jars. you are just generating steam with it. or four, four full liters of autoclave... heh.

So you just steralised the jars...not the agar ? I thought u were meant to steralise the agar in the jars at the same time and steralise both ?

@fresnomycologysociety5004

3 жыл бұрын

I sterilized the agar with jars and then filled them. It's not too difficult. The problem with sterilizing the agar in the jars is that you have to make sure the agar is homogenized, which requires it to boil a bit. For that reason I just let it mix in the PC.

6:38 - forbidden pudding 😬

God originally tried this method for human procreation but was like, "Fuck it. No one's gonna buy a pressure cooker. Just let 'em fuck'n figure it out."

You could have prefilled your jars with agar before sterilization so you didn’t have to pour your Petri dishes in the SAB.

you might as well do No Pour TEK

What can you substitute for the malt extract?

@activeenglishlecreusot9628

5 жыл бұрын

clear honey ;)

At 1:19 I thought tom petty's mary janes last dance was starting

ooof just buy the pre made mix yo...

where can you get agar powder at?

@fresnomycologysociety5004

7 жыл бұрын

www.amazon.com/gp/aw/d/B008SV7P8I/ref=yo_ii_img?ie=UTF8&psc=1 this is the agar used specifically in this video. this 16oz bag will make this 500ml recipe 50 times. which is alot. You can call your local health food stores and Asians markets too, they would be selling it as a vegan substitute for gelatin.

@rockstar0777.

7 жыл бұрын

+Fresno Mycology Society thanks a lot!!

Why doesn't everyone just put the jar on the scale and zero it before adding the next ingredients? It sure would save time and cleaning of measuring containers!!!

Or you could just do no pour tek since you poured into small mason jars lol

9g agar. 10g light malt extract. 500 ml distilled water.

@lilkaps3344

5 жыл бұрын

Light malt extract powder right?

@FreshPotential

5 жыл бұрын

@@lilkaps3344 Correct, powder.

@Mk-to1ln

4 жыл бұрын

Do I need distilled water

your using your bare skin on the surfaces there, even with IPA that'll screw things up. i like this kinda rustic method of mycology though

@paladro

6 жыл бұрын

certainly using proper equipment would be the preferred method, but if it works, how can you argue? i'd take a flow hood over a glovebox, but a glovebox over open air. rustic or 'ghetto' methods can work, but you should expect mixed results.

A good proposal for your sterilization box îs to attach a 8W UV lamp - under 20 dollars from AliExpress -has small dimensions and can be disconect before pooring Agar. Many bad comments below will be solved, regarding sterilization.

@63phillip

3 жыл бұрын

That would have to be a UVC light and it would have to be kept away from all living cultures. but good to sterilize equipment and box or even substrate before Inoculation.

FIRE UP THE PC

For please in spañish mat. No speak english good job exelent

Music to loud

agario

Glove box is a huge nono

Why are you calling this a substrate, its NOT a substrate.

@thinginground5179

3 жыл бұрын

Why not?

music or dialog never both ok.

My pinset is still bigger than yours

llol i though this is agar.io -_-

@naomimarquez9451

7 жыл бұрын

same

@cosmitmixes7048

6 жыл бұрын

KING/ كنج same

This would be so much better without the "music"