Needle biopsy for cloning fruit to liquid culture. Lots of tips for sterility and aseptic technique.
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80% of the time it works 100% of the time
Hey Ed was just listening to one of your older lives while cloning with your needle biopsy techniques 😂 maybe I had to start a bit later
This is exactly the video I have searching for flaws and all. We all are going to have issues so its nice to see how one can recover from them. Thanks for this.
Yes Sir, right i On time.
Thanks for all the information and what you do for the community 🍄❤️. I quit smoking and I vape now but when I do work I tape my vape to something so i can hit it without my hands 😅
I don't actually vape while I work but in between jobs, I have a small work area so I can only do a few things and then I have to re set up but I keep it off to the side I don't breathe all over my work station or anything. 😅
Yes I better should do it after this post. I didn't clear the needle and it was difficult to get some tissue into the needle. Luckily after some trials I flame sterilized the needle and now I know why it worked better. Thanks Ed for the great info, keep it up, you make my myco life a lot more successfully.
I just realized I didn’t put any lme in my distilled water. I’m such a new goober
Stop smokin' so much weed man ;)
Just got my order, thanks my man, keep on keeping on🍄
I love it. Don't stop.
This guys a real Gem !! Thanks Ed !!!
I've been cloning a lot of gourmets from the Asian supermarket. I saw your first video on this, but I never tried it. I'm going to give it a go this weekend!
Hydraulic effect has screwed me more than once and in many different fields.
Thank you ❤
Never used needle for cloning myself but I think you might have a better chance if you twist the syringe couple of times while going through the stem.
I forgot where I first seen it,but I spray(sanitize) the air too, lol
I'm guessing oust would be the best for that
I keep getting contam when I spawn to bulk so would it be better if I use bags?
If your spawn is good, try boiling/sterilizing your substrate. Coir is dirtier than people think.
@@edwardgrand I bought it from mycohaus online and it was supposed to be ready to go but I’m going to make my own this time and use bag’s
Good video, I get it
I used my imagination thanks for this, can you show progress on the jar in future? That would be cool and scientific.
I keep forgetting to ask in Lives but have you much experience in the bunson burners cone of sterility and can you work plates and tissues in the cone with good results or is a still air box better?
When I work in a stab I feel like its definitely going to contam.
Thanks again for taking your time to teach.
I have worked with many SABs, burners, flow hoods, FFUs, etc. All of it comes down to technique rather than actual equipment. Being aware of how contams spread and what NOT to touch, etc. is the key.
I'm thinking best to take a biopsy length ways with the fibres of the fruit. Rather than trying to cut through them cross ways?
Why did you throw the cap on the ground 😂 thats good eaten man. I think people get overly ocd about being sterile.
My alcohol is denatured. Extremely bitter to eat.
@edwardgrand you took the cap off before spraying it I believe 😄
@@devilsadvocate1338 I have enough ;)
@@edwardgrand 🤣 I bet
Hey doc! Thanks for another incredibly informative video! I didn't know simply spraying down the mushroom was enough to ensure sterility for an LC inoculation! I usually split the stipe and use the needle to scoop a piece from tissue inside. I also never knew the cone of sterlity was so big! And sucking through the flame to get sterile air is genius!!
To be extra safe, what do you think of palcing the mushroom in alcohol, and then performing the tissue biopsy while the mushroom is submerged in the alcohol? I figure this way, the tissue will be in blocking the needle from drawing up an appreciable amount of alcohol and there should be zero risk of any surface contaminants making their way into the tissue sample or needle tip.
Finally, would H2O2 be adequate to surface sterilize for this process or is alcohol still better?
You're welcome. I don't think submerging the mushroom would be necessary or practical. Alcohol seems sufficient for surface sterilization, but I reckon peroxide might be good, too. I don't like spraying too much peroxide. It tends to bleach everything.
@@edwardgrand Ah but do you think it would reduce the chance of contamination, and more importantly -- would the tissue still be viable? I'm concerned about surface contaminants making their way into the LC, especially if the mushroom was taken from a less-than-ideal environment. Or if I happen to be too engrossed to notice my kitten sneezing nearby. Or there is an undetected draft in the room, etc.
My understanding is that introducing even a *single* endospore will ruin the entire jar of LC, so I would prefer to minimize the chance of contamination, however small it may be. Especially because contaminants in LC can only be identified by plating after the LC has colonized, I personally prefer to reduce the chance of contamination to as close to zero as possible.
Btw, your portable bunsen burner is so awesome! Idk if this is at all practical but with a larger mushroom, could you skewer it and do a real quick roast to char the outside surface instead of spraying disinfectant? Or maybe even spray the disinfectant and flambe the mushroom briefly!
@@vincentwu2848I haven't had trouble with contam and this technique.
@@vincentwu2848Even if you do all that, you still won't know without plating it.
@@vincentwu2848I can't answer all your questions in one go. I just wrote a 500 word response and accidentally deleted it.
Fruits can be extremely blue without any interference, even at firm or recently soft harvest time. Happens to me all the time. I have never sprayed them with iso haha
Love you brother, danke
Thanks Doc 💎⚡️
Thank you Ed!
What genetics are the cluster your working with? 🤘🤘🤘
Melmac derivative
Where you get them lids?
I made them. It's in another video.
th-cam.com/video/_9dbAa5T0g4/w-d-xo.htmlsi=tzv97CLPO5Bkvm0v
What is the recipe or what amount of dry malt extract for 500ml of liquid culture, thanks for any info and thanks for the awesome video 👍
0.5% = 2.5 gram
@@edwardgrand thanks 🤟
@@edwardgrand mushluv fungifam 🤟🍄🤘
pro tip to open up sterile packa of syringes and needle tips: wet the paper side with alcohol and then push the locking parts through the paper and quickly lock the two pieces. after that you can take your time and safety pull them out of the packaging. you can do this in less than a second and no need to fiddle around