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3.A.M
Boop!



Registered: 10/17/22
Posts: 847
Loc: Oz
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Multispore streaking with gill fragment 3
#28607883 - 01/03/24 12:26 AM (24 days, 22 hours ago) |
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Decided to put my neck on the block and officially put this little technique I’ve been practising out there for the shroomery crew to critique, shoot down or hopefully advance further.
The basic theory is to dissect a small piece of gill from as close to the cap as possible thereby avoiding possible contaminates that accrue through the process of a grow then streaking the gill over agar to produce a clean ms grow. I’ve successfully attempted this 4-5 times now and am hoping it will be extremely helpful in the process of refining phenos in my cross attempts, I’ll outline the process I’ve developed below which is stupidly simple but I’d suggest making sure your cloning skills are up to scratch beforehand as it’s essentially the same process with a slightly different end game.
What I do first is tear from the base of the stipe upwards so that it also splits the cap and you have 2 halves of the same shroom to play with, I then cut diagonally downward starting from just inside the cap to about 1/2 way or less if possible into the gills, then the same again at an opposite angle creating a V shape, from there I cut side to side through the cap so I have a little triangle of gills and a piece of cap to reinforce and stabilise it, this is extremely important as without it the piece of gill will disintegrate . Here’s a basic diagram of the process
 Now when I make that final cut through the cap I angle it a little downwards towards the gills, from there it’s a matter of working with what you’ve got, I’ve had the piece simply lift up with a single gill attached, sometimes a couple which doesn’t matter, sometimes I have to slide my blade under the gill and lever it up so it tears where I made the incision in the cap but once it’s separated make sure it doesn’t make contact with any part of the shroom where it can potentially pick up cross contamination, then I simply stab my blade through the cap section marked below in red.
 From there I drag the gill lightly over the agar (I make my agar fairly soft so there’s more chance of spores sticking) use your common sense and drag away from the sharp edge of the scalpel so you don’t cut through the tissue mid drag or save yourself potential trouble altogether and sterilise some tweezers and use that instead, again making sure to grip the cap tissue only. Here’s a plate I streaked a few days ago, no pattern, I simply use the gill like a brush painting back and forth over the centre of the plate.
 Not the best image but you get the idea. Yes, there’s definitely varieties and species this simply won’t be practical for, most small pans for example but I’d love to hear peoples feedback, positive or negative, it all helps further our general knowledge and skills but I really hope some of you give it a try and let me know how you go, Cheers folks!
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inski
Cortinariologist



Registered: 02/28/06
Posts: 5,720
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Re: Multispore streaking with gill fragment [Re: 3.A.M] 1
#28607892 - 01/03/24 12:45 AM (24 days, 21 hours ago) |
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I've done it quite a few times over the years and it certainly works but is more prone to contamination compared to using a properly prepared spore print. It's also possible that part of the culture you get starts from hyphae from the gill fragment, in essence a clone which is not a problem.
It's definitely possible to acquire a viable culture with this method.
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3.A.M
Boop!



Registered: 10/17/22
Posts: 847
Loc: Oz
Last seen: 6 hours, 21 minutes
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Re: Multispore streaking with gill fragment [Re: inski] 1
#28607899 - 01/03/24 01:11 AM (24 days, 21 hours ago) |
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I was hoping someone else had tried this and would respond, I’ve definitely had fragments clone on the plate but they were usually pretty obvious, one grow I couldn’t be certain if it was ms but I haven’t had contamination yet thankfully, but I can’t remember the last time I got contam from cloning either, could be luck or technique it probably helps that everything I’ve tried it with so far had fairly large, dense gills which probably makes a huge difference also.
Edited by 3.A.M (01/03/24 02:57 AM)
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LogicaL Chaos
Ascension Energy & Alien UFOs




Registered: 05/12/07
Posts: 69,298
Loc: The Inexpressible...
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Re: Multispore streaking using a gill fragment [Re: 3.A.M] 1
#28607924 - 01/03/24 01:39 AM (24 days, 20 hours ago) |
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I like it!
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