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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: Pasty Agar Tek [Re: taGyo]
#20936569 - 12/06/14 07:45 AM (9 years, 1 month ago) |
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Have a panellus stipticus (bioluminescent bitter oyster) culture received a couple of days ago. It's only a 3rd of an LC culture and I don't need to pour a whole sleeve of petris to expand upon it, so I am going to make up a few of some no-pour plates (5) to expand this culture out with. The method I use is not exactly Pasty's style, but the same principles apply. Just want to drop by and give a shout out to the no-pour method of doing agar for when you don't need to pour a whole sleeve of petri dishes at a time to expand upon a small amount of culture right away.
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Pasty Agar Tek [Re: hamloaf]
#20936580 - 12/06/14 07:51 AM (9 years, 1 month ago) |
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when you go LC>agar, do you move the syringe into the agar at all? or just noc the surface of the agar?
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Huh, usually just the surface. Never heard of moving the syringe into the agar to inoculate agar with LC. That's a new one. I am going to have to try that.
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Pasty Agar Tek [Re: hamloaf]
#20936600 - 12/06/14 07:59 AM (9 years, 1 month ago) |
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I meant like creating a little valley in the agar with the syringe for the LC to pool up in, so its not just all over the plate. that way you can control the growth... I mean, that is at least my thinking. Have yet to do it.
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Pudgy
Fail-2-Win

Registered: 09/30/14
Posts: 657
Last seen: 8 years, 9 months
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That's a pretty neat idea. Even for MS.
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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I see what you mean. My mind went over a few possibilites when I first read that statement.
That actually strikes me as a good idea as a possible preventative measure from keeping the culture from also sliding all over the surface of the agar when freshly inoculated. I'm going to have to try it on a few of these panellus stipticus inoculations. I will report back here.
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Pudgy
Fail-2-Win

Registered: 09/30/14
Posts: 657
Last seen: 8 years, 9 months
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Re: Pasty Agar Tek [Re: hamloaf]
#20936618 - 12/06/14 08:06 AM (9 years, 1 month ago) |
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I'm kind of curious what would happen if you did inject just under the surface of the agar. Would it crawl out?
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Re: Pasty Agar Tek [Re: Pudgy]
#20936637 - 12/06/14 08:11 AM (9 years, 1 month ago) |
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yeah it would grow up through the agar, back onto the surface
agar sandwhich isolation tek has proof: http://www.shroomery.org/8513/Agar-sandwich-isolation-TEK
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,918
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I've seen people do that..I forget who it was but someone actually recommended it.
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taGyo
Strainiac/AMU



Registered: 10/16/14
Posts: 18,802
Loc: Journal Land
Last seen: 5 years, 3 months
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Re: Pasty Agar Tek [Re: hamloaf]
#20936656 - 12/06/14 08:18 AM (9 years, 1 month ago) |
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Quote:
hamloaf said:
That actually strikes me as a good idea as a possible preventative measure from keeping the culture from also sliding all over the surface of the agar when freshly inoculated. I'm going to have to try it on a few of these panellus stipticus inoculations. I will report back here.
PLEASE let me know. The first time I did LC > Agar this happened:

Whoops, I meant this:

Looks like I inoculated with a shotgun but it was only a drop or two that must have spread out. It turned into this:

Which was really annoying to sector. I had a nice piece of rhizo growing in there but by the time I prepped my agar the next day it had moved into a tomentose area. This is KSSS which I find on agar has a tendency to be big and puffy. I heard this is a sign of multiple genetics?
-------------------- Gyo's Better Grows TNF Q&A AMU Q&A Dominus fortunae meae sum
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,918
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Re: Pasty Agar Tek [Re: taGyo]
#20936672 - 12/06/14 08:24 AM (9 years, 1 month ago) |
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The tubs in my sig were all tomentose. I must put too many nutes in my agar or something and rarely see rhizo growth..but it doesn't effect much IMO. There are still multiple genetics there yeah...not sure if you can confirm that though by it being "big and puffy", but you'll need more than 1 transfer or so to get a monoculture.
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taGyo
Strainiac/AMU



Registered: 10/16/14
Posts: 18,802
Loc: Journal Land
Last seen: 5 years, 3 months
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I think Pasty was the one who looked at 'em and said I had a lot of genetics because it wasn't flattening out :/,
All my transfers are growing well so I'll just keep sectoring
-------------------- Gyo's Better Grows TNF Q&A AMU Q&A Dominus fortunae meae sum
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Imperfect Iam
^means imperfect,not I'm perfect



Registered: 03/05/13
Posts: 7,237
Loc: center of the universe
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Re: Pasty Agar Tek [Re: taGyo]
#20936743 - 12/06/14 08:51 AM (9 years, 1 month ago) |
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An even faster way to get a monoculture is to grow out a transfer, then take a clone, instead of going throught possibly 100's of Petris to get your monoculture.
I am not so sure it would take 100's, but I know of some very experienced cultivators on here, who would tell you to take a clone, maybe they will chime in on that one.
-------------------- All you touch, and all you see, is all your life will ever be- Pink Floyd Life is what happens while you're busy making other plans- John Lennon
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taGyo
Strainiac/AMU



Registered: 10/16/14
Posts: 18,802
Loc: Journal Land
Last seen: 5 years, 3 months
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So grow out my agar plate and then inoculate and grow that out for a better chance at a mono? I also agree that going through 6-7 plates sounds ridiculous .
-------------------- Gyo's Better Grows TNF Q&A AMU Q&A Dominus fortunae meae sum
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Imperfect Iam
^means imperfect,not I'm perfect



Registered: 03/05/13
Posts: 7,237
Loc: center of the universe
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Re: Pasty Agar Tek [Re: taGyo]
#20937497 - 12/06/14 12:06 PM (9 years, 1 month ago) |
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Exactly, grow out a culture, then take a clone to get your monoculture. A lot more fun than repeatedly transferring cultures, without ever seeing a fruit, IMO!
-------------------- All you touch, and all you see, is all your life will ever be- Pink Floyd Life is what happens while you're busy making other plans- John Lennon
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MrGiraffe

Registered: 04/04/14
Posts: 3,149
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Even if you make a transfer and then fruit it, a cloned fruit from that fruiting can still have multiple sectors. I had a pink buffalo clone that had more sectors than I cared to count. I fruited it out, and took another tissue sample and it still has a shit ton of sectors. I then isolated some of those sectors and surprise, even taking tiny tips of rhizo growth from these plates that I thought were single sectors, grew out into more sectors. I did however use this process to end up with a mono culture of a nepal fruit though, so it can work. If nothing else by cloning a fruit, you at least know you have a fruiting culture.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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I don't isolate my clones down to mono cultures. I find they preform just fine as they are most of the time.
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MrGiraffe

Registered: 04/04/14
Posts: 3,149
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Re: Pasty Agar Tek [Re: MudaFuka]
#20937625 - 12/06/14 12:45 PM (9 years, 1 month ago) |
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But how do you know what your putting into or taking out of a slant each time you start a new plate if there are multiple substrains within the slant? I remember readin PussyFart's A Tale of 10 Isolates. He started with tissue clones of 3 separate fruits, then isolated out. Some of the isolates were pure shit. Without isolating, how are you sure you don't end up with one of those on the plate you're growing out?
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taGyo
Strainiac/AMU



Registered: 10/16/14
Posts: 18,802
Loc: Journal Land
Last seen: 5 years, 3 months
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Thanks for the interesting read Giraffe, Never seen that thread before. Makes me not even want to mono-culture tbh
-------------------- Gyo's Better Grows TNF Q&A AMU Q&A Dominus fortunae meae sum
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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That's why I don't isolate my clones. I keep them intact so I know how they will preform. I don't make slants and when I inoculate I use the whole plate. My only problem arrive when I transfer to expand to new plates. There is always the chance of creating an isolate inadvertently. I could eliminate this by using LI made from a whole plate to inoculate several new dishes instead of just transferring small pieces of mycelium. I think that's what I'm going to do from now on thanks for making me think about that one.
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