Spore Print to Agar - The easiest and most efficient way to use and clean-up spore prints.
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- Опубліковано 26 вер 2024
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Edward Grand UA-cam: / @edwardgrand
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Your chuckles are freaking amazing and legit made my day lol, thanks for sharing. I'm doing this today!
I just did my 1st spore to agar may 3rd and now have growth today I'm so happy
I was actually going to ask you if there was a better way than an ennoculation loop and now I don't have to because here it is. Makes so much sense and preserves your precious prints. Fantastic! Keep em coming brother!
What's the issue with an inoculation loop? It's vastly more simple than this technique
@doloinc because when you try to drop spores onto a plate in front of a flow hood they usually blow away..... it's really difficult to get them to land on the plate
@@trevorbraden5448 I've never seen any issues spores blowing away. Ever. It sounds like inoculation technique issues
@doloinc I lose alot of spores in front of my hood..... some still land on the agar..... but a lot get waisted..... blowing past the plate...... spores don't weigh anything so doesn't take much for them to blow around
@doloinc if you don't need a loop that's one less piece of equipment to buy and one less piece of equipment to sterilise.
I've been using a cool little dental pick with a ball on the end instead of a point. Used for some kind of cavity filling. I just dab the spores with the ball end and used the flat end to spread them. It's actually probably using too few spores. I'm going to try this method from now on. Thanks!
Man this video was right on time I've been struggling with placing spore prints on plates for awhile now.... they always blow away in front of a flow hood......This is genius....
Your videos are my favorite way to start the morning!
good stuff indeed
Another excellent and educational video. Coming from a physical science/engineering background I've found you're a valuable resource as I am picking up experience in mycology. I'm backing out of a liquid culture that has at least yielded opportunity to make spore prints and doing no pour agar later today. Now, if I only had some color shift plasti-dip pearls to go along with the light malt extract ;)
Thanks Ed...great stuff as always. Also, it is so good hearing you say things like "i have no idea what it is...we'll see" when you are working with contaminated plates and such. Thanks!
just brilliant with the agar. learn something new every time tnx ED
WoW this technique is just awesome ❤
Thank you for sharing that !!!
another great video brother, keep it up, youre good for the community
I’m so glad you created this video. I’m going to be do some agar projects and this will be the tek I use ty Ed!
I like this, I also like the inoculation loops for this. I used your rip swab tek last night.
Thanks Ed. You are my hero
Just received some prints , heading to sab now !
Thanks for video !!
Another great tip that I will use next time. Thanks, Ed!
Hey Ed! what are you working on today? Hope you are doing GREAT!!! 🍄❤
Nice one bro I appreciate the time an work yall put n I love learning new things.
Tweezers eye surgeon uses would be awesome for this. Super fine point have 3 pairs now have a new use for them besides digging out splinters lol
Awesome, I will definitely try this out when I get some wild prints!
Nice I gotta try this!!!
Very cool man I like the way you make it simple . So when you grab just the little cotton hair for xfer, are you ever thinking that just that tiny little piece, vs a chunk, will be a great narrowing of the gene pool? Or would the fact that it's maybe still a T1 or T2 mitigate that?
The idea is to narrow down the gene pool so you can fruit something consistent. It really depends on what it looks like on the new plate. Could be ready for spawn, but some people like variety. The main idea is to make sure it's clean. A smaller piece will more likely eliminate competition from other mono- and dikaryons and possible contaminants that may be lurking.
Thanks Ed
Oh wow that looks so much simpler
Dang! Wicked technique!
Very nice! Thank you sir!
I cut the a little aluminum and drop it in a dropper bottle but I like this better
Nice one Ed. Cheers
Nice video fam
My partner was just asking me about this the other day! Word.
Great video! Are you doing this all in front of a hood? I wasnt sure if it's a good idea to do spore and early culture stuff in front of a hood because im worried about contams getting blown all over my work place. And since this stage seems highly likely for contams, just wanted to get others opinions.
Whoa! I've never seen this technique anywhere else. I've seen people recommend swiping spore swabs in a Z shape across the plate. What's the reason for the Z-shape swipe, as opposed to your technique of touching the spores to three places on the plate?
Also, is your laminar flow hood delivering sterile air from the top? Do you have any thoughts on which type (vertical or horizontal) are better suited for mycology? Thanks!
The idea of 'streaking', as it's called in microbiology, is to get single spore (or bacterium) separated from possible contaminates and other spores. It was also the impetus behind me doing the 'grab and drag' technique. With the grab and drag, the idea is to get a single spore released from the swab cotton tuft, which then germinates, and can be carefully isolated and hopefully will develop into a monokaryotic colony that can be used for mating. In bacteria, streaking can be used as a rough estimate of CFU (colony forming units), but serial dilution is a much more precise way to do that. In fungi breeding, all we need is one monokaryon to proceed, so we don't need to be too concerned with how we get it as long as it's a monokaryon.
Re: laminar flow hoods
It doesn't matter if it is vertical or horizontal. I have worked with both and it is simply personal preference and individual work flow. If you are doing most mycology stuff (e.g. grain, bags, etc.) I would suggest a horizontal flow FFU. I frequently use a vertical type, but that is because of some logistical issues, not really my preference. TBH, a still air box (SAB) is fine, but when you get to a certain volume of work, it becomes awkward and impractical. I've actually done a fair amount of work using the 'cone of sterility' techniques using a Bunsen burner and alcohol wiped desk. I wouldn't suggest it for long-term, but you can get away with it sometimes (isolated away from contams is one time).
Wow!
Ed I love your videos BUT.... Only joking. I have a question. I made myself a methylated spirits burner with a hemp wick. It doesn't burn very hot. I went back to using my map gas torch. However, I notice that your burner isn't very ferocious either. Does the item that Iam trying to sterilise need to be glowing red hot? Other mycology UA-camrs often say to get them red hot. Thanks for your time 🖤
You can listen for the scorching sound when you touch it to agar. If it makes a sound, it is more than hot enough. I also let them rest in 70% alcohol between uses. You can use a cigarette lighter in a pinch. Between transfers, you're really only trying to kill the last thing it touched, which hopefully is clean mycelium, so you don't need to get it crazy hot.
@@edwardgrand thanks for the answer Ed. I really appreciate it. Last time I used the torch I ended up with black deposits (I don't think it was soot) off of the scalpel. I'm guessing I over did it that time, or these scalpels are a sub standard material
@@edwardgrand I will keep a small jar of iso on hand for that purpose👌
@@kevinfromdevon Soot is sterile. The fungus doesn't mind. Some people put carbon in their media. :)
@@edwardgrand I'm pretty sure that the black deposits were burnt myc or agar. Doesn't appear to have affected anything and I've since transferred away from those dishes again. Thanks for the reply Ed
I tried this technique on 4 different prints and just ended up with bacteria growing on my agar. I'm guessing the prints are not clean enough to start with.
Yes. I hate to say it, but spore printing requires good technique. I have seen things that send shivers up my spine...
@@edwardgrand I think I will keep using the inoculation loops until I am working with my own prints. Later today I will be trying out your tweezer method on some swabs that just came in the mail
@Kevinfromdevon if the prints are dirty, it might take some proper streaking technique to isolate clean spores. It sucks. Adds weeks to the effort.
@@edwardgrand I received them from experienced cultivators, so Iam hopeful that I can get something from them. I can only cross my fingers at this point and see what I can do. Thanks for the replies ✌️
Wouldn’t that contaminate the spore if the agar was bad? Id scrape spores onto agar.
I usually pour my plates a week before using them. No reason they would be contaminated. The idea is to separate the spores from possible contams on the print by 'streaking' them across the plate.
Where are you located? Interested in meeting to share techniques?
I'm far away from the Western world. Except this computer terminal. LOL
@@edwardgrand I live in the san francisco bay area. I am enjoying your videos about breeding.
Do you have a preferred agar recipe for spore germination?
I have been using this for everything:
SRYA recipe
3% Sticky Rice powder/flour
2% Agar
0.1% Baker's or Nutritional Yeast
Food Coloring (if desired)
FOR 500 ml Total volume
15 g sticky rice powder
10 g Agar
0.5 g Baker's Yeast
Food Coloring or 1g charcoal if desired.
500 ml tap water (not distilled or DI)
- Should make between 20-25 standard size plates.
But 2 % MEA works fine, too. There is no magic recipe. If you have trouble germinating on MEA, it is likely the spore's problem, not the media.
Not Really ! even a little bit agar left of the print will be enough to ruin that print. Tricky, Not Recommendd !
I make sure the agar spot is dry before packing it back up. Seems fine.
It wouldn't ruin the print at all. A tiny bit of agar, in dry conditions, won't be enough for the mycelium to really start growing.
Excellent content. Follows the KISS method to the T (K eep I t S imple S tupid)👍