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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
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Re: So I hear it's hard to print PE? [Re: mushpunx]
    #22329930 - 10/03/15 07:35 PM (8 years, 3 months ago)

I would say you shouldn't do LC without something to sterilize it.

If you get septum-top bottles though, it's pretty easy to be sterile once you can PC the media.

If you need sterile spores you could always try fruiting it in a bag...

Does PE not produce them at all, or just very few?

Very few is okay but then you would certainly want agar since inoculating substrate with that would be painfully slow.


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InvisibleFantastic Mr. Fox
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Registered: 02/12/15
Posts: 3,225
Loc: Flag
Re: So I hear it's hard to print PE? [Re: YaMoonSun]
    #22329957 - 10/03/15 07:42 PM (8 years, 3 months ago)

Just because you see growth doesn't mean it's clean growth.


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InvisibleGrey
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Re: So I hear it's hard to print PE? [Re: micro]
    #22329967 - 10/03/15 07:45 PM (8 years, 3 months ago)

They produce them, they just don't drop them.


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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
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Re: So I hear it's hard to print PE? [Re: Fantastic Mr. Fox]
    #22330263 - 10/03/15 08:57 PM (8 years, 3 months ago)

Amen, but I'd like to take my chances. Worst possible outcome? :shrug:


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OfflinecronicrFacebook
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
    #22330417 - 10/03/15 09:38 PM (8 years, 3 months ago)

Quote:

YaMoonSun said:
If I can't print it, I should still be able to empty a syringe of sterile water into a cap and quickly stir it up and pull it back into the syringe, no? Bad idea? Ever try it? Likely need to get a SAB for that procedure. :shrug:



it has worked out before, i'd expect a little failure along the way though(should have a sab for everything)


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Offlineinvitro

Registered: 05/03/13
Posts: 2,529
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
    #22330675 - 10/03/15 10:40 PM (8 years, 3 months ago)

:thumbup: good info Grey, thx


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InvisibleViolet
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Registered: 12/06/11
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Re: So I hear it's hard to print PE? [Re: invitro]
    #22331287 - 10/04/15 03:25 AM (8 years, 3 months ago)

That's cool, about the centrifuge.

I have used the syringe drops method to harvest PE spores before into a syringe.  I found it more tedious than using a swab but equally, possibly more, rewarding.  Thing is the droplet's surface tension stops it from easily getting down into the gills. And when it does, it's not coming back out.
So what I did was:
1. Sterilized a syringe with 5-8 mL of water in it, with the needle attached, all wrapped in foil.
2. Used my extra mature and wrinkly PE fruit that had grown invitro - picked in my sterile airflow work space, and readied the syringe in my other hand.
3. Placed a few droplets of water on the gills while holding the fruit upside down.
4. Worked the water into the gills with the tip of the needle. Repeated 3 and 4 until excess water that could easily be sucked up was building up in the cap.
5. Without absolutely shredding the gills, ran the needle tip along the gills, with the goal to rub off spores into the water.
6. Sucked as much of the water up as I could.

If you feel good about having this all kept clean during the process, which I did in my sterile airflow, you can shake the syringe and put water back in to stir it with the water that remained from the first time and try to get more of the spores that you freed but were left behind.


To me, this was worth it. Because with 1mL of spore solution I can start like 10 of my grain petris for my culture tech.


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OfflineYaMoonSun
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Registered: 10/23/14
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Re: So I hear it's hard to print PE? [Re: Violet]
    #22331992 - 10/04/15 10:05 AM (8 years, 3 months ago)

:takingnotes:


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