How to Make Agar Plates for Growing Mushrooms
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- Опубликовано: 9 сен 2024
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Agar plates are also used for making fungal clones. A clean fragment of a mushroom fruiting body can be placed on agar to grow rather than germinating spores. This gives the cultivator and advantage in isolating any potential contaminants if present.
Recipe : 750 ml Hot water + 15 Grams Light Malt Extract + 15 Grams of Agar powder.
Recommended Links:
Light Malt Extract Powder: amzn.to/3d2cRkY
Agar Powder: amzn.to/3BreXnV
Sterile Petri Dishes: amzn.to/3QBxAda
Para Film: amzn.to/3cYhzA1
Laminar Flow Hood Parts:
HEPA Filter amzn.to/3BZWTl2
Blower Motor amzn.to/3QWmcso
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I have just decided to move away from growboxes to start my own personal ''farm''. And your videos has been extremly helpfull :) Thanks for upload m8!
I’ve really been thinking the same thing ! Lmk how your adventure has been !
@stixgrim8064 this is also my goal...are there other RUclips channels that you also like and are helpful?
I use boroscillicate petris , reusable as long as they don't break. I also use 599 ml Pyrex media bottles (all autoclavable )
Amazing video, very easily explained and feel very confident in my first attempt with agar . Great job 👏 👍 💯
Love your Tito's bottle.
Love your videos! Thank you so m7ch for making them.
🙏🏽❤️🙏🏽
@@jakedominguez-street Thanks for watching!
Awesome vid deserves more views
I appreciate that! 🙏
Fantastic job and thank you so much for sharing
Thanks for that great video..
Thanks for watching!
Great video!
Great vid thanks. So condensation on the plates don’t matter? Isn’t it better to leave the plates in front of the hood without lids until they solidify ? Or would that increase chance of contamination? I have a flow hood so I was thinking of placing them right in front
Yeah you should cover the plates as soon as possible, you can store your plates upside down and the condensation will not fall on to the agar/mycelium.
Great video! Why should I use agar plates and not spore syringes what are the advantages and disadvantages of each method?
You should use both. They are both different from one another. Agar plates are a great tool to germinate the spores from the syringe and isolate individual strains from it. Agar plates are also useful for longer term storage of the mycelium.
Agar plates ensure your not using a contaminated spore syringe before you waste all your substrate by injecting it with contaminated syringe
i see that many people use nutritional yeast with this method
does it do anything? should i just leave it out
Hey bro what’s the foil for inside the lining of the PC? I have the presto 16 and 23 quart so I’m curious if I’m missing something ? Also,is that parafilm or grafting tape that you used? I have a box of parafilm now but was using a roll of cling wrap (also gas exchangeable) and cut the roll into 1-2 inch pieces and worked with that. Micropore tape always leaves a residue on the dishes which can allow contams to stick easily. Mush love!
Sometimes my fruiting bags would melt if touching the sides, didn’t happen anymore after I lined it with tinfoil. Some people use wet towels, idk maybe I was pressure cooking higher than 15 psi on accident and that’s why it would melt when touching the sides. Either way, doesn’t hurt.
Great stuff. Thanks
How do i make it without a pressure cooker?
Do you have a video or images of your flow hood? I saw the links below of what to buy but curious how you set it up? Also do you have a hammock filter on it as well?
Yes I have images but I can’t post them here, I’ve built a couple of them. You just build a box around the filter leaving space behind the filter and then attach the blower on top above that space. You can also build a box around the blower motor and add the pre-filter to that.
If you want to email me at Sporensprout@gmail.com I can try to help if you run into problems.
What was the hole in the Tito’s bottle cap for? You never did anything with that.
It just provides gas exchange. If you don’t have a hole in the cap, it will build up pressure.
@@SporenSprout oooh I see. Thanks!
Sterilized it in the pressure cooker
Great video. When you wrap your dishes with condensation how do you get that to go away?
You can put a heating pad on top of the dish to help get rid of the condensation. To make sure there is no condensation to begin with make sure to pour at 125 F. If you pour hotter than 125F there will be condensation.
hey, great video. quick question: what is the suitable method for storing them? Also, for how Long can you store them before use?
Store them on a shelf at room temperature. Temperature fluctuation draws out moisture from the agar and causes it to dry up around 6 months. Keep at steady temperature for 6-12 months and transfer to new plate before it drys up.
Just asking, do you loosen the lid to your bottle before you sterilize? otherwise it seems like the bottle would blow up from the pressure.
Yes it also has a small hole with tape over it.
What's your method for getting rid of all that condensation? That's my biggest problem yet with agar. Plates are flawless crystal clear, but by the time they've sat in the sab and cooled, nutrient plates all get spots. Any advise?
Yeah just pour when the agar is cooled down enough for you to touch with your hand and there won’t be any condensation.
Rack the the plates one above the other
Heat from above plate will remove condensation on below
I've seen several people selling charcoal agar plates. Would be pretty cool to watch the mycelium grow against a black background. Do you think itd be safe to mix a little activated coconut charcoal into this recipe?
Yes it is. Drop me a message if you need a good recipe, with peptone.
@@MrJosh-bd9rpI need one please
Can I use a crock pot instead of a pressure cooker 😅
You can try steam bath for 2 hours. You will know if it failed after one week because there will be contamination growing on the plate. If nothing grows after a week then it’s ok to inoculate.
Great vid. Can I do the pouring in a fume hood? Will that be sufficient to avoid contamination?
Thanks! I would not work around anything but either completely STILL air or hepa filtered air. It’s worth a shot though.
@@SporenSprout I wipe down the inside and outside of the hood with 91% alcohol before I do any bacterial inoculations on purchased pre-pours and get good results. I'm gonna try this protocol with your method...I'll let you know how it turns out.
@@goodboysic Should definitely work! Good luck!
Another great video I'm going to grab a bottle of Tito's as soon as the liquor store opens it's a great excuse to get myself some good vodka for a change! Is your flow box homemade?
Thanks for your support! 🙏 I had a feeling people will go out and buy some Tito’s just for this 😂 Yes my flow hood is homemade!
Any chance you might do a video on how you made it? How much do you have invested in materials?
@@christianlander321 Yeah I was planning on making a video soon. Cost around $550 for 24” x 24” which is the perfect size.
Looking forward to seeing that video for sure! I'm a carpenter by trade so the box construction is no biggie I'm just curious about the HEPA filters and blower motor specs
@@christianlander321 Novatek F4010 HEPA Filter Novair... www.amazon.com/dp/B01EC60IG8?ref=ppx_pop_mob_ap_share
Dayton 1TDT2 Blower, 549 CFM,... www.amazon.com/dp/B000BK859C?ref=ppx_pop_mob_ap_share
Simple and nice process, but what are these things that look like bubbles on top of plates..?
maybe condensation from the heat from the agar mix?
Can I just use Nutrient Agar? what is the function of malt extract? Greetings and excellent video
You can use nutrient agar by it’s self. The light malt extract is used for nutrients when using regular agar agar, which does not have nutrients and will not support healthy growth on its own.
@@SporenSprout Great, thanks for responding, I'll probably use potato water instead of that
How long can they keep without mycelium on them? Best to keep them in the fridge? I’ve bought a pack of twenty but I only need 2 currently. Guessing it’s best to do then all now and store them rather than bag them back up and chance contam?Thanks spore ✌🏻
They actually keep for quite a long time especially if there isn’t a lot of temp fluctuations drawing moisture out of the agar. Usually still good to use up to 6 months at room temp.
What to do if theres lots of sedinent on the bottom? It doesnt dissolve completely
Make sure the water is hot and mix it again
Myc will eat it
In another video he added yeast to it too, i couldn't understand the logic behind it , since we don't want to culture yeast or any other organizm but mushroom on it, then why yeast, so i came to this one.
No I never added yeast to agar. It was a broth for cordyceps and it was nutritional yeast not activated yeast.
@@SporenSprout thank you
Hello. In my country it does not allow me to buy: "Agar powder" from Amazon because the audana does not allow it, what other implement can I use?
I’m sure they have agar powder at a local grocery store. It’s a common ingredient in food.
Where can I buy Agar already made and what does it do?
Bruh
hey man what kinda of pc is that?
Presto 23 quart.
Does it have to be light malt extract would I get away with ordinary malt extract
Yeah that will still work, Here is the updated video though: My Favorite Recipe for Making Agar Plates Full Tutorial
ruclips.net/video/otBU3jlUZAE/видео.html
Hi I can only find the syrup like malt extract it has a honey consistency can I use this still ?
Could you put the mixture directly into the Mason jar and pressure cook it like that?
Yes that works well.
100% That's the only way I've previously done it.
sorry but I have another question, what are the proportions that I should use for 10 petri dishes?
it´s around 300ml of water, 6g of agar and 6g of potato water am I correct?
You need to give up trying to grow mushrooms. 300ml for 10 plates. So 30 + ml's per plate. So you're trying to fill them to overflowing, where do you put a sample. The linear volume of a standard 90mm x 15mm plate is like 22ml. Soooooo, you can't make plates and your comment about agar and spore syringes was absolute nonsense. Stop it dude
Your number is 312ml total. 10 petrie dishes have a total volume of 225. Soooooo you just pour that 87 extra ml's on the floor or drink it or what. Where'd you get this math?
I did a trial of 2 agar plates. One didn’t work the other one did and it is full of mycelium. What do I do now?
Store the plate in the cold or transfer wedges to new plates and expand some wedges to LC media and sterilized grain.
What temperature they do they need be stored ?
Good at room temp for 6 months and over a year in refrigeration.
What kind of pressure cooker do you use? How many quarts is it?!
I have a 41 quart electric all American but I used to use a 23 quart presto which was fine.
Quick question. How long can you store agar before transferring mycelium to it?
Until the agar starts drying up which will happen after 6-12 months.
Do you store them in the fridge or just room temp?
Is that roll of parafilm considered parafilm "grafting" tape??
Yes I prefer it.
why do you use a bottle and not a mason jar? can you use a mason jar that isn't full?
Yes a mason jar works, the long neck makes it easier to pour.
First of all, mason jars pour like shit, and an empty liquor bottle is just whatever is around. How about you get yourself a couple borosilicate media bottles, 500ml, with drip lip. Pressure cook with cap sealed no problem 30 min at 15psi. Stop being cheap, and stop for the love of God asking ignorant non scientific questions. This dude said why use agar and not spore syringes and the advantages etc.. bro!!! Not even the same anything to compare. Agar is a tool, spore syringes are meant to be used in combo with said tool not one or the other. Just sounded dumb, I'm sorry. Most of you should just buy shrooms, lol.
so he just poured the mix in there
After sterilizing the liquid agar it came out with only 1/3 of the contents inside the bottle! I did everything as you said.. what did i do wrong?
Loosen the lid before pressure cooking. Make the hole in the lid a little bigger. Don’t go over 15 psi. Make sure there’s enough water in the pressure cooker.
Thanks!! But why would that be? I thought I made the hole too big (wouldn't that be why the water escaped in the first place?), if I loosen even more, won't the water come out more?@@SporenSprout
I see some people just using the agar and malt , why do some ppl use potato flakes and corn syrup ?
You can use a lot of different stuff to make agar plates. You can even use strained grain water.
Sometimes you need just water agar with no nutrients for the purpose of isolating individual spores for breeding. It just depends what is easily available to you and what your goals are.
@@SporenSprout just bought agar powder and LME . I’m good to go with that ?
@@Allyourbase1990 Yeah you’re good
I have agar but not nutrient rich should I try using some sugar in this recipe?
My agar is the telephone brand
Honestly you only need light malt extract and any type of agar. You would have to mess with the measurements but yes you can replace light malt extract with grain water, honey, potato water, or basically anything really.
Where did you get the big roll of para film? :)
amzn.to/44hgitm
That bottle trick tho..
Can I substitute the malt extract with any other ingredient or take it out of the recipe?
The malt extract provides nutrients. If you take it out you will have water agar plates which are generally used in single spore isolation techniques to slow down growth. You can substitute malt extract with a bunch of other stuff.
@@SporenSprout, could you please tell me some of the most common stuff to substitute the malt extract? It's hard for me to get it where I live. Thanks in advance.
@@JorgeMartinez-bruy potato water, grain water, dextrose, nutritional yeast.
Basically you’d substitute water for grain water or potato water.
@@SporenSprout, I really appreciate your explanation!
How do u store the plates and how long will they last in storage
I store them on a shelf at room temp while they grow and once they are mostly colonized I put them in a mini fridge. Usually good for 12 months or longer like that. If left at room temp they will start to dry up after 6 months.
If you need longer storage, here’s the vid for culture slants:
ruclips.net/video/MSL9sYez-CM/видео.html
@@SporenSprout once the agar mix sets would it be bad to store them in the mini fridge at a low temp setting rather than a shelf at room temp?
@@SporenSprout thanks for the link man i needed it
@@smokintuffllc1496 That would be fine to do.
Hi what is the substitute of malt extract?
Potatoe flakes or beer yeast
what are the storage conditions for these plates? both when they contain spores and when they do not?
Just keep them on a shelf in a room that has a steady room temperature.
Isolab autoclavable pitchers. You’re welcome
Do the autoclavable pitchers come full of liquor though? They should for the price of them.
why do you use alcohol bottle? can we use a regular jar instead?
I like the long neck. Yeah you can use a jar. Here is another video you can watch about making agar:
ruclips.net/video/otBU3jlUZAE/видео.html
Can u use a rigid plastic bottle??
Yeah but it has to be PP5 plastic. I use glass quart jars with one air vent in the lid and those work too.
@@SporenSprout thanks