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Sivarted
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Registered: 08/22/15
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Re: Odds on getting anything viable out of this syringe? [Re: Inocuole]
#22434627 - 10/26/15 03:23 AM (8 years, 3 months ago) |
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And I was all worried and shit...
4 pasty plates, all of them showing nothing but white. 
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Sivarted
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22434629 - 10/26/15 03:25 AM (8 years, 3 months ago) |
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Knocked up a couple quarts of wbs too, and there's one tiny little spot of growth already, even though it's only been a few days since I did that.
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LocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ



Registered: 04/17/15
Posts: 7,076
Loc: to the brain
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22434632 - 10/26/15 03:28 AM (8 years, 3 months ago) |
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Q&A US vs. THEM The more I learn, the less I know.
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22435070 - 10/26/15 08:44 AM (8 years, 3 months ago) |
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Quote:
Sivarted said: And I was all worried and shit...
4 pasty plates, all of them showing nothing but white. 

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Sivarted
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Re: Odds on getting anything viable out of this syringe? [Re: wowimflabbergasted]
#22442892 - 10/28/15 12:33 AM (8 years, 3 months ago) |
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And...Definitely contaminated. First two plates I did were good, and looked like this:

Second two, a few days later, were not. I did just one or two drops on the first two, next pair I did one drop at a time spread around, and got a couple spots of white but mostly little brown volcano looking things.

And one if the quart jars I knocked up looked like it had a spot of myc growing, but that disappeared and now the only thing that's showing up is a gooey brownish stuff.

But! Hope is not lost. Got two transfers from one of the first two plates, one from the other, and a tiny myc scraping from the center of one of the volcano filled ones, so I'm still pretty sure I can clean it up. 
Oh yeah, and that brown glob I managed to squirting out has regenerated. Going to hang on to the syringe until I get something clean just in case, because there are definitely spores in there, but as soon as I have clean cultures that sucker is going away.
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22442920 - 10/28/15 12:46 AM (8 years, 3 months ago) |
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APE can be a bitch. All my syringes are nothing but bacteria but it's not because the looks of the spores.
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Sivarted
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Re: Odds on getting anything viable out of this syringe? [Re: wowimflabbergasted]
#22442951 - 10/28/15 01:03 AM (8 years, 3 months ago) |
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Quote:
wowimflabbergasted said: APE can be a bitch. All my syringes are nothing but bacteria but it's not because the looks of the spores.
A bit of digging around does suggest those dark circular patches are bacterial colonies. Looks like I'll be able to get something out of it though, so that's good.
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22442956 - 10/28/15 01:06 AM (8 years, 3 months ago) |
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Can't see that well, but it looks like bacteria to me, perhaps it was under the mycellium to start with, then the mycellium died due to lack of access to nutes under it? IDK
Molds usually look like less structured mycellium. That brown stuff does not look like sporulation.
Make the vendor make that syringe right. I'd just grab the culture you have and get some different more basic syringe. PE6 sounds like a worthwhile cube strain.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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Sivarted
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22442972 - 10/28/15 01:13 AM (8 years, 3 months ago) |
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The interesting part of this is that both times I used the syringe on agar, the first couple drops were clean, or at least had more spores than bacteria. The first two plates, I think I only put one drop each on, and they both have myc growing on them. The second pair, the first plate I did had two spots of myc in the center, where I dropped first, and every drop after those first couple appears to be bacterial.
I did reflame before the second plate, but didn't reshake the syringe, so I wonder if the shaking forced the spores toward the needle for a short time and the bacteria to the other end. If that's true, I should at least be able to get more agar samples. I might play around with that idea and try to get a few more.
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22442989 - 10/28/15 01:21 AM (8 years, 3 months ago) |
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I might try gently dragging a needle over some fluffy mycellium if you have any, grabbing a small amount, then dragging it over a new plate.
It could be the vendor sold you a LC (PE syringes being difficult to make well,) which is getting more bacterial over time. That might explain that weird looking glob in the middle too. IDK.
I wouldn't over analyze it too much. Maybe you just got lucky using less solution.
The only active measure you could take that I can think of is holding the syringe vertically needle downward. Spores seem to be very heavy, that might concerntrate them above the needle. I think they must be more dense than bacteria, since ?Workman? said they centrifuge ground PE caps to make the syringes, and the spores are the bottom layer. Again, I'd only use one drop. Asside from reducing the chances of putting a contam on the plate, you're making less of a liquidy mess on the plate, and something that can evaporate more easily. Getting solution all over is probably a great way to spread bacteria.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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Sivarted
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Registered: 08/22/15
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Re: Odds on getting anything viable out of this syringe? [Re: Machiavelliavore]
#22571092 - 11/25/15 12:54 AM (8 years, 2 months ago) |
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Oh hey, looks like I might get one quart of spawn from that after all.

Thats been going for almost a month from MS inoc, and the myc will probably be weak as shit, but it might overrun the bacteria and give me a little tray worth of ape. I'd be happy with even a handful of pretty blue alien fruits. 
Haven't been keeping up on the agar transfers very well in my quest for getting a bunch of isolates to test though. Just did my second round of transfers, bringing the total to 16, but the transfer was about a week late, so it's not super likely I pulled any isolates out.
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Sivarted
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
#22571098 - 11/25/15 12:58 AM (8 years, 2 months ago) |
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The vendor made good and gave me credit btw. Since I've got a bit of agar growing (9 plates that are 4 sections each) I'll probably pick up a different strain with it, but they did come through and try to make it right.
Pretty sure I'm not supposed to mention the name even though I'd like to, because I have clearly grown out the spores, but it was a sponsored site. 
I decided to try to get the original two clean pasty plates to pin so I could clone a pin or two as well, opened them up and put them in my FC, but it's not looking promising so far. We'll see.
Edited by Sivarted (11/25/15 01:02 AM)
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