How to Make Agar Slants for Mushroom Cultures EASIEST METHOD
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- เผยแพร่เมื่อ 20 มิ.ย. 2024
- This video gives a walkthrough on how to make agar slants for preserving mushroom cultures. Agar slants are great for storing fungal cultures long-term. Culture slants can last months or years with no maintenance! Slants are much better than agar plates for long term storage of mushroom cultures because the air tight screw-cap prevents the culture from drying out over time. The easy method presented here uses a media dispensing bottle to rapidly fill many tubes without a mess! Filling each tube with a syringe or flask can be tedious and messy. Using the plastic dispensing bottle allows to you to seamlessly fill many tubes.
Media recipe:
500ml water
10g agar
10-12g potato dextrose or malt extract
Additional media recipes and more:
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Chapters:
0:00 Needed Supplies
0:24 Agar Prep
0:51 Filling the Culture Tubes
2:39 Cooling the Slants
3:51 Inoculation
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More info on how to grow mushrooms at home! th-cam.com/video/W3xsqxZBnQ8/w-d-xo.htmlsi=sjMmYgIcm7CbRhDc
Nice clear instructions, thanks
Thank you sir.
Thanks for watching 🍄
Are there pros/cons to using a scalpel vs. inoculation loop? Great tutorial! Thanks!
The inoculation loop is a little easier for me to use when working with slants. The only downside is that you're not going to get as much tissue with the loop. When working with a scalpel you can cut out a large piece of tissue if you need and it'll colonize a little faster. But keep in mind that you might want to preserve your master slant as long as possible so taking a smaller piece of mycelium with the loop could be more advantageous in many instances.
Quality info
Glad it was helpful!
You got a link for that scalpel? I love how slender it is. Makes slant work easier and I bet it cools faster after flame sterilization than what I'm currently working with.
Found it, looks like it might be a #7?
Yup, you got it
Thank you for your videos! They are very educational.
I am more familiar with your old videos. Trying to catch up now. I feel my knowledge is a bit rusty after over a year break from studying mycology. I had experimented little and due to health conditions I was forced to have a break and my LC spoiled meanwhile.
I’m ready to restart but now all I have is 1 spore print and 5ml spores suspended on water. Unfortunately I am not able to get more.
I have materials for grains on jars, bags, agar, LC, bulk substrate… What would you recommend I start with so I make the best of the little I have? For grains I have access to rye and popcorn by the way. I am interested in starting a good self sustaining cycle loop so I don’t waste the spores I have.
Your help would be greatly appreciated. Thank you in advance!
Hi, thanks so much for watching! I would recommend starting simple with just a bag/jar of rye grain spawn. Use 1-2ml of your spore solution to inoculate it. You can then use that grain spawn to inoculate 5-10 more bags/jars of grain. I think this is the best method for working with a limited amount of inoculum. Once you have grown those bags to completion you can then make some fresh LC, fresh plates and more to start buking up your inoculation material.
Really appreciate your comment! Let us know how it goes! 🍄
@@MotherMycelium thank you! I will try that and hopefully I won’t have any contaminants. Do grains last longer than LC then? Or is just because it produces more?
@@thiagorossi3860 LC should last much much longer than grain. I'd recommend starting with some grain and growing it all the way out to completion so that you can make your LC with a clean tissue clone.
@@thiagorossi3860 If you're just trying to make LC you could put a few drops of your spore solution on agar and then use that agar to make LC once you've confirmed it's contaminant free
Wooww, don't mind me stalking your page lol but how do you exactly get it out once it's colonized? Does it just squeeze out?
Once it's colonized you can use the methods that we used to inoculate the slant. You can simply cut a small piece of mycelium out of the colonized slant with a scalpel or inoculation loop. Then you can keep the slant in storage and use the freshly transferred mycelium.
Should the slants be colonized after the transfer at room temp, or should they go right into the fridge and can colonize over time at 40 degrees?
We always let them colonize for a day or two and then refrigerate. This way you can check for contamination and such.
Does it matter if you use potato dextrose or LME? Would it make a difference in terms of shelf life of the slants?
Any agar nutrient will work here. To ensure a long shelf life, seal with a few layers of parafilm and store in a plastic bag. Some people also add sawdust to the solution or a popsicle stick help with the shelf life.
@@MotherMycelium Thanks bro, mush love ❤️
I like this method however I found condensation inside the slants. Next-time, what do I do differently to negate the condensation?
If you are having issues with excess condensation you can try letting the agar cool a little more before closing the lids completely. You can also pour the slants and then leave them open and let them cool in front of a flow but I generally don't recommend that unless it is totally necessary.
why do you add agar to the slant before inoculating it with the fungal inoculum?
You need to wait until the agar is completely cooled and solidified before you inoculate it. I like to heat up the agar mixture to dissolve it, then add the mixture to the tubes and sterilize. Then you let the slants cool completely before you inoculate them. If you incoulate the agar while it's still hot and liquid you will kill the fungus
I don’t need nearly this many slants. What would be the ratio for day like 5 of these?
Thanks for watching! If you're just making enough agar for 5 slants you'll need about 50ml (10ml per slant). You'll need about 50ml water, 1g agar, 1g malt extract.
However, I would recommend that if you're gonna go through the process of making slants then you should just make a larger batch and store the extras for later use. 🍄🌍
Is there a reason why 20 minutes @15psi instead of the standard 90 minutes @15psi that grain jars/bags are sterilized?
Liquids generally take a lot less time to get up to temp. 15-20 min at @ 15 psi is typical for sterilizing liquids with autoclaves.
You also want to avoid caramelizing the sugars in your liquid culture/agar solutions
How come they always boil over? Any help
I would recommend filling your slants with a little less agar or using slightly larger slants.
I guess I missed that part where the condensation is removed.
I like this method however I found condensation inside the slants. Next-time, what do I do differently to negate the condensation?
Aww man, i don't have a lab coat 😔
Can you place the slants directly in the pressure canner without anything like jars or racks?
You could but you'll need a way to keep them upright so the agar solution doesn't spill out. Keeping them in the tube rack or mason jar also helps to easily remove the tubes after they are sterilized
@@MotherMycelium dude thanks for the quick response! 🙏🏽🤘🏽