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OfflineSivarted
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Odds on getting anything viable out of this syringe?
    #22400891 - 10/19/15 12:34 AM (8 years, 3 months ago)



APE syringe from a sponsored vendor.  Presumably it has spores in it as well as whatever the hell is growing in it, I'm going to try to get something on agar, but I'm obviously not super hopeful.  oO

Probably just going to end up waiting for a replacement.


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InvisibleLocN9ne
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22400902 - 10/19/15 12:44 AM (8 years, 3 months ago)

What's wrong with it?


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OfflineSivarted
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Re: Odds on getting anything viable out of this syringe? [Re: LocN9ne]
    #22400945 - 10/19/15 01:15 AM (8 years, 3 months ago)

Quote:

LocN9ne said:
What's wrong with it?




Other than the huge growth of what looks like some kind of mold in it?  Nothing I guess.  Definitely does not look like a spore clump though.  :smile:

And from what I've read, APE spores are supposed to be clear, and practically invisible in water.


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OfflineMachiavelliavore
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22400989 - 10/19/15 01:41 AM (8 years, 3 months ago)

Yeah that looks pretty bad.  If you can take a single drop and put it on agar, then get the spore company to send a replacement.  Whatever that huge clump is, if it's living it's probably gonna stomp all over some cube spores.

Shit, that even fucks it up for microscopy :smile:

Maybe inject some into a roommate and see if you get the fluke man from the xfiles.


Oh yeah, they make those PE syringes by centrifuging ground caps, maybe some other crap from htat process got sucked in.  It might be inert.


--------------------


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...


Edited by Machiavelliavore (10/19/15 01:41 AM)


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Invisiblewowimflabbergasted
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Re: Odds on getting anything viable out of this syringe? [Re: Machiavelliavore]
    #22401657 - 10/19/15 09:00 AM (8 years, 3 months ago)

Try it and see dude the APE syringes that I had all had white shit in them not black spores. Not sure why I never grew them.


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OfflineSivarted
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Re: Odds on getting anything viable out of this syringe? [Re: wowimflabbergasted]
    #22403065 - 10/19/15 11:10 AM (8 years, 3 months ago)

APE doesn't have black spores from what I've read, they're transparent.  I dunno.  Whatever that glob is, it's not spores.  I know what spore clumps look like.  Still going to give it a shot.  Knocked up a brf jar and a couple agar plates, we'll see.


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OfflineSivarted
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22418021 - 10/22/15 12:15 PM (8 years, 3 months ago)

Update from the vendor:

Quote:

Thanks for writing.

The sample you received should be OK. The spore material used for APE is not dark purple/black like most of Ps. cubensis... it is much lighter, lacking the pigments. So when the spore clusters clump up like this it can be a bit nasty looking. If you find the material to be contaminated after performing microscopy, please let us know!




Not really convinced, because I've seen spore clusters before, and that does *not* look like a spore cluster (plus it's a really nasty looking brown), but since APE doesn't actually drop spores maybe it's fine and that's just material that goes in the syringe with the spores?  I dunno.  We'll see.

I inoculated one brf jar and two agar plates, the agar this morning just started showing tiny dots of what might be myc, so hopefully even if the syringe is as contaminated as it looks, I can at least do some transfers on it. :smile:


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Invisiblewowimflabbergasted
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22419633 - 10/22/15 06:49 PM (8 years, 3 months ago)

Its normal. APE syringes aren't going to look like other ones you've seen.


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InvisibleMad Season
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Re: Odds on getting anything viable out of this syringe? [Re: wowimflabbergasted]
    #22419700 - 10/22/15 07:01 PM (8 years, 3 months ago)

:whathesaid: looks like a syringe from a certain vendor with a centrifuge.. :wink:. All the syringes I've gotten from them have huge globs of spores in the syringe like that. I'd bet all in that that is a great ape spore syringe right there.

Actually.. Its bad. Send it to me to make sure :wink:


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Re: Odds on getting anything viable out of this syringe? [Re: Mad Season]
    #22419708 - 10/22/15 07:02 PM (8 years, 3 months ago)

:lol:


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Re: Odds on getting anything viable out of this syringe? [Re: wowimflabbergasted]
    #22420793 - 10/22/15 10:51 PM (8 years, 3 months ago)

Don't assume that syringe is bad. It does look funky but you can't judge them visibly.
Try 2-3 jars.

Had that syringe been used before?


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OfflineMajickMuffin
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Re: Odds on getting anything viable out of this syringe? [Re: tahoe]
    #22420818 - 10/22/15 10:56 PM (8 years, 3 months ago)

Looks like a liquid culture .

What if somehow the spores have germinated and are forming a mycelium network inside there. Company made mistake ?


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InvisibleMad Season
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Re: Odds on getting anything viable out of this syringe? [Re: MajickMuffin]
    #22420828 - 10/22/15 10:57 PM (8 years, 3 months ago)

It looks like albino spores to me lol. Albinoism effects spores too. This is why aa+ isnt a true albino. But leucistic A+ isn't anywhere near as cool haha


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Edited by Mad Season (10/22/15 10:58 PM)


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Re: Odds on getting anything viable out of this syringe? [Re: Mad Season]
    #22420923 - 10/22/15 11:25 PM (8 years, 3 months ago)

My APE syringe germinated while being stored in the fridge. Looks quite similar. Just tested it and have good growth.


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OfflineSivarted
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Re: Odds on getting anything viable out of this syringe? [Re: Pastywhyte]
    #22421309 - 10/23/15 02:41 AM (8 years, 3 months ago)

I have tests underway.  That big clump got stuck in the needle and the whole thing shot out at once though.  Presumably there are still spores in the syringe though.  Oops.  :smile:

The agar plates I did showed their first signs of white though, so it seems like everything is good.


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InvisibleInocuole
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22421350 - 10/23/15 03:30 AM (8 years, 3 months ago)

All that not being convinced for what? :lol:


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OfflineLogicaL ChaosM
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Re: Odds on getting anything viable out of this syringe? [Re: Sivarted]
    #22421748 - 10/23/15 07:34 AM (8 years, 3 months ago)

That sure is strange. Never seen that before.

Then again, ive never grown an albino strain either :shrug:


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Invisiblewowimflabbergasted
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Re: Odds on getting anything viable out of this syringe? [Re: Inocuole]
    #22421755 - 10/23/15 07:38 AM (8 years, 3 months ago)

Quote:

Inocuole said:
All that not being convinced for what? :lol:



:whathesaid::lmafo:


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Re: Odds on getting anything viable out of this syringe? [Re: MajickMuffin]
    #22427920 - 10/24/15 03:53 PM (8 years, 3 months ago)

Quote:

MajickMuffin said:
What if somehow the spores have germinated and are forming a mycelium network inside there. Company made mistake ?


I have read that the spores can cannibalise their own tissues. With poor spore droppers I expect gills might be in the spore solution.

I have gotten exotics with visible myc growing in them, never cubes, and I have gotten far more cube syringes over the years.


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Re: Odds on getting anything viable out of this syringe? [Re: blackout]
    #22427932 - 10/24/15 03:55 PM (8 years, 3 months ago)

It would suck if that's what grew out instead of the spores, and it happened to be from a clone culture near the end of its life.  With something like APE that's actually somewhat likely to be the case, at least comparatively to other varieties.  Hopefully some spores would germinate and integrate into the genetics and keep that from being an issue.


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OfflineSivarted
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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)

And I was all worried and shit...

4 pasty plates, all of them showing nothing but white.  :smile:



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OfflineSivarted
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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)

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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InvisibleLocN9ne
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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)

:vibin:


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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)

Quote:

Sivarted said:
And I was all worried and shit...

4 pasty plates, all of them showing nothing but white.  :smile:





:lmafo:


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OfflineSivarted
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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)

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.  :smile:

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.  :smile:


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Invisiblewowimflabbergasted
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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)

APE can be a bitch. All my syringes are nothing but bacteria :lol: but it's not because the looks of the spores.


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OfflineSivarted
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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)

Quote:

wowimflabbergasted said:
APE can be a bitch. All my syringes are nothing but bacteria :lol: 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. :smile:


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OfflineMachiavelliavore
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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)

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.


--------------------


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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OfflineSivarted
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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)

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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OfflineMachiavelliavore
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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)

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.


--------------------


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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OfflineSivarted
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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)

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.  :laugh:

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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OfflineSivarted
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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)

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.  :smile:

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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