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Offlinepoisoned
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Getting tissue off the scalpel
    #26552296 - 03/23/20 04:06 AM (3 years, 10 months ago)

It's all fun and games when you're doing a plate to plate transfer, where you cut out a nice square and just pick it up with the knife. But I'm having a shit ton of issues when making clones and when transferring from slants. Especially with ganoderma. I bought a culture, but getting the mycelium out of that bottle is already a PITA. Then making it stick to agar again is an even bigger PITA. It just wraps around the scalpel and won't get off. Then it makes me all nervous and I contaminate my plates when I wrestle with the piece of myc. I always try to use a fresh blade, so the surface of the scalpel is smoother, but it's still giving me problems.

Any suggestions on how to solve the problem?


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OfflineHaywire
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Re: Getting tissue off the scalpel [Re: poisoned] * 2
    #26552426 - 03/23/20 06:21 AM (3 years, 10 months ago)

when cloning mushrooms, you can use a pair of tweezers to pull some tissue from the core of the stem.
What I usually do is I cut across the width of the mushroom about 1 cm apart. then you can also pick some tissue from the stipe.

if the agar wedge is sticking to the blade, just cut into the receiving plate to dislodge it.
when you take the wedge from the donor plate, just take it with the tip of the scalpel. So it's just barely hanging on.
If that doesn't work, you can increase the strength of the agar. more solid agar wedges are easier to pick up and place in the receiving plate.

It just takes some practice. a good way to do this, is practice outside of the still air box with non inoculated plates.
just cut, pick and inoculate. do this fifty times and you'll see there's really nothing to it.
It's the stress of not wanting to fuck up a good plate that makes it difficult.

Good luck.


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Offlinepoisoned
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Re: Getting tissue off the scalpel [Re: Haywire]
    #26552510 - 03/23/20 07:19 AM (3 years, 10 months ago)

Yeah, plate to plate is easy. I have issues with slants and clones. With slants, it's more like with clone, where you can't really cut out the square, you have to scrape the myc and transfer it. Stabbing the receiving plate doesn't really seem to help with that. I think i need to improve my scraping technique.


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InvisibleLadysKnight
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Re: Getting tissue off the scalpel [Re: poisoned] * 1
    #26552589 - 03/23/20 08:03 AM (3 years, 10 months ago)

I've found when transferring a scrape, it comes off easier if I cut into the receiving plate backwards from how I took the scrape. So if I scrape with the blade down, I transfer with the blade upside down.


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Offlinepoisoned
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Re: Getting tissue off the scalpel [Re: LadysKnight]
    #26552967 - 03/23/20 11:55 AM (3 years, 10 months ago)

Sounds like a good idea, I'll try that!


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OfflineHaywire
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Re: Getting tissue off the scalpel [Re: poisoned] * 1
    #26554496 - 03/24/20 06:24 AM (3 years, 10 months ago)

I use a loop to transfer from slants. just scrape it across the surface and then on the receiving plate.
it takes a little longer to establish but it works.


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