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OfflineFivel
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Bacterial syringe
    #20578942 - 09/17/14 10:00 AM (10 years, 3 months ago)

Hello guys. I need a little advice. I put some spore solution from a vendor syringe of Penis Envy on agar less than 48 hours ago. Both plates I did appear to be overrun with bacteria. I am wondering what I should do at this point? Is the syringe just done? Or will I be able to transfer myc from the plate once it germinates(if it germinates :sad:)
That's they only thing I can think of at this point is to wait for it to germinate, transfer it. and then wait for it to grow to a clean spot on the new plate because there isn't any way I will be able to take a clean sample from these first 2 plates..


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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InvisibleMr. Alien
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Re: Bacterial syringe [Re: Fivel]
    #20578954 - 09/17/14 10:03 AM (10 years, 3 months ago)

Agar is the solution for your problems. You can isolate away healthy myc from contams with agar, you can easily germinate your spores there, clean it and guarantee a contam free culture.

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OfflineFivel
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Re: Bacterial syringe [Re: Mr. Alien]
    #20578960 - 09/17/14 10:04 AM (10 years, 3 months ago)

I start everything on agar.. That's where they are now lol. My prob is the whole plate is overrun with bacteria and I have no germination yet. The syringe was a little iffy.. It came out kind of gloopy if that makes sense..


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

Edited by Fivel (09/17/14 10:05 AM)

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OfflineSh33p.Milk
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Re: Bacterial syringe [Re: Fivel]
    #20578963 - 09/17/14 10:05 AM (10 years, 3 months ago)

I've had success transferring a small amount of a clean colony from a contaminated plate.  Getting a clean colony can take a few transfers, but if there's hope, don't give up.
Vendor syringes can be delivered pre disturbed, so there is a chance that your sterile technique wasn't at fault.
Keep updating and add pics!
Good luck

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InvisibleMr. Alien
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Re: Bacterial syringe [Re: Fivel]
    #20579015 - 09/17/14 10:23 AM (10 years, 3 months ago)

Sorry i read to fast your first post.

Ass pictures, there must be a piece of myc at least.

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OfflineFivel
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Re: Bacterial syringe [Re: Mr. Alien]
    #20579031 - 09/17/14 10:30 AM (10 years, 3 months ago)

lol there isn't. The spores have barely had time to germinate. They have been on agar less than 48 hours. The bacteria has grown over 90% of both plates.. I guess I will just wait to see if I get germination. If they germinate I'm pretty confident I can save them but it's going to take some transfers..


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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OfflineSh33p.Milk
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Re: Bacterial syringe [Re: Fivel]
    #20579088 - 09/17/14 10:51 AM (10 years, 3 months ago)

And the anxious waiting game begins! If it does work and you make it to your next dish, you could consider using anti-bacterial agar.  I know Fungi Perfecti sells it.

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OfflineFivel
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Re: Bacterial syringe [Re: Sh33p.Milk]
    #20579102 - 09/17/14 10:58 AM (10 years, 3 months ago)

Holee shite. Good idea.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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InvisibleMr. Alien
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Registered: 01/14/14
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Re: Bacterial syringe [Re: Sh33p.Milk]
    #20579117 - 09/17/14 11:01 AM (10 years, 3 months ago)

Quote:

Sh33p.Milk said:
And the anxious waiting game begins! If it does work and you make it to your next dish, you could consider using anti-bacterial agar.  I know Fungi Perfecti sells it.




:whathesaid:

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OfflineFivel
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Re: Bacterial syringe [Re: Mr. Alien]
    #20579135 - 09/17/14 11:08 AM (10 years, 3 months ago)

Gentamycin Sulfate Antibiotic Powder would work don't you think? It would help if nothing else right?


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Offlinedeathlyego
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Re: Bacterial syringe [Re: Fivel]
    #20579181 - 09/17/14 11:23 AM (10 years, 3 months ago)

What do you guys think of the sandwich tech? (once he had mycelium growth)

http://www.shroomery.org/8513/Agar-sandwich-isolation-TEK

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Offlinespacechildo
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Re: Bacterial syringe [Re: Fivel]
    #20579183 - 09/17/14 11:23 AM (10 years, 3 months ago)

sounds really un-neccessary.
how much spore solution did you squirt on the agar?

from a syringe you can squirt on your loop and then swipe the loop on agar.
if these are poured to petris are you sure your pouring tech is clean enough?

and as always you really need pics, no one describes stuff the same way.

sandwich tek > antibac agar!

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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Bacterial syringe [Re: Fivel]
    #20579226 - 09/17/14 11:39 AM (10 years, 3 months ago)

I had this issue myself not long ago. I tried inoculating a single grain jar and see what happened, as I never had contamination going this route, but my agar contaminated from a select few syringes to the point where it took over the plate before mycelium was even visible. Ended up working for the most part, though I havent gotten around to expanding that jar via gtg.

But try inoculating a grain jar, from there you can take a few grains and put those on agar. I think thats what I'll do. But the jar did end up fully colonizing.

Edit: Here is my post http://www.shroomery.org/forums/showflat.php/Number/20346667

Edited by Hyphai (09/17/14 11:41 AM)

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OfflineFivel
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Re: Bacterial syringe [Re: Hyphai]
    #20579259 - 09/17/14 11:47 AM (10 years, 3 months ago)

I'll nocc up a grain jar with it then. I don't have a good enough camera for pictures. There really isn't much to see. Huge bacterial blotch all over the plate with a small black spot of spores in the middle. I used 2 drops of spore solution per plate. Will squirting it on a loop and then putting it to agar do anything different? I assume it will be a little better cause I am getting rid of a lot of the H20 from the solution. I ordered the Gentamycin anyway. If I end up not needing it now at least i'll have it on hand for later use. :shrug:

Also, I do no pour. So no worries about it being sterile before I open it. I do all my work in an SAB. I inoculated 4 other plates with 1 other syringe and a print at the same time I did these two. The other 4 look clean and doing what they are supposed to be doing I assume. This one just has shitty bacteria all over it  :dead:


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

Edited by Fivel (09/17/14 11:53 AM)

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Offlinespacechildo
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Re: Bacterial syringe [Re: Fivel]
    #20579334 - 09/17/14 12:03 PM (10 years, 3 months ago)

Quote:

Fivel said:
Will squirting it on a loop and then putting it to agar do anything different? I assume it will be a little better cause I am getting rid of a lot of the H20 from the solution.




:yesnod:

are there satelite colonies of bacteria or large splotches all over?
it can be a real bitch so if you have other plates that are fine just use them and discard the baccy one!

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OfflineFivel
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Re: Bacterial syringe [Re: spacechildo]
    #20579362 - 09/17/14 12:09 PM (10 years, 3 months ago)

Large splotches all over. Also, let me tell you what I have going. I put 3 different, eh.... species (I think.. I don't wanna get wrapped up in variety/species debate) to agar. I used 2 plates for each. So I have 2 P. Galindoi Plates from print. I have 2 Pan Trop plates from syringe. I have 2 P. Cubensis Penis Envy plates from syringe. The 2 Penis Envy plates are both covered in bacteria. The other 4 are fine. No growth whatsoever.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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