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OfflineCelestialexplorer1
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Uv-c to sterilize agar * 1
    #26643426 - 05/02/20 10:49 AM (3 years, 9 months ago)

Ok so I know there are topics in uv-c light for sterilization and a lot of people are against it but I had a thought. Since with agar what you are doing is working with the surface of it, and uv-c is used to sterilize surfaces and found rather inexpensively on Amazon.. well I think you get the idea.

Just wondering what others might think of this idea. Obviously I would still pc the agar and use all sterile techniques but just to go a step further you could make a chamber with the light to hit the surface of the agar to kill any possibly lingering bacteria.


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Invisibleninja cat 09
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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26643450 - 05/02/20 11:00 AM (3 years, 9 months ago)

It's an interesting idea, I think it would be great to sterilize a glove box or workspace in front of a flow hood, but I don't know what it would do to the glove box plastic, plus, I understand that you need eye protection to be around those types of lights.


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OfflineCelestialexplorer1
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Re: Uv-c to sterilize agar [Re: ninja cat 09]
    #26643635 - 05/02/20 12:46 PM (3 years, 9 months ago)

Well with what I’m thinking it would be more like a glovebox or trunk that is solid in color and non-transparent with a switch on the outside. So you put your agar in (after the cook and pour), close the lid tight so no light can come out and flip the switch and then turn the switch off BEFORE opening to ensure safety. I’ve seen uv-c bar lights on amazon for 30$


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Invisiblenooneman
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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26643640 - 05/02/20 12:50 PM (3 years, 9 months ago)

It might work if you do it for long enough. The problem with UV is not that it's ineffective, it's that it causes cancer. So like the previous poster said, if you built an enclosed chamber with no light escaping, and let it bake under UV for a couple hours, it couldn't hurt. Just make sure you limit your exposure.


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OfflineCelestialexplorer1
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Re: Uv-c to sterilize agar [Re: nooneman]
    #26643762 - 05/02/20 01:38 PM (3 years, 9 months ago)

Google search revealed up to 8 feet it will make organisms inactive in 30 minutes. So that’s not very long and in a non transparent glovebox and with no peeking it should be safe. Probably do some more research though before deciding to try anything.


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Invisibleninja cat 09
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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26643974 - 05/02/20 03:09 PM (3 years, 9 months ago)

Quote:

Celestialexplorer1 said:
Google search revealed up to 8 feet it will make organisms inactive in 30 minutes. So that’s not very long and in a non transparent glovebox and with no peeking it should be safe. Probably do some more research though before deciding to try anything.



You could make agar by boiling it, not PC the agar and then put plates of it in the UV-C chamber for varying amounts of time to see what happens.

I remember a tech twitter person I follow, who is reputable (@RealSexyCyborg) mentioning that some LEDs that are sold as UV-C aren't actually UV-C, so you'll probably want a way to test them before you buy something off amazon.


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Offlinekitten6
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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26645300 - 05/03/20 06:26 AM (3 years, 9 months ago)

well if your lamp is powerful enough, it can actually turn oxygen in air into ozone which will do a pretty good job of sterilization, it will kill rats as well. Your room will stay completely sterile for a good while. As long as it smells a bit of ozone.



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Re: Uv-c to sterilize agar [Re: kitten6]
    #26645318 - 05/03/20 06:41 AM (3 years, 9 months ago)

Don't worry about discoloring everything in the room eventually.


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Invisiblemycorry
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Re: Uv-c to sterilize agar [Re: bodhisatta] * 1
    #26645806 - 05/03/20 11:48 AM (3 years, 9 months ago)

Last time I did research on this I discovered that almost everything in agar(including the antibiotics if you use them) will be damaged or destroyed by the UV-C.

Furthermore, I let a set of plates that I had sterilized with UVC sit in their ziplock bags for a few days, and upon opening them they all smelled like ozone. I don't want a little cloud of ozone hanging out near the surface of my agar ready to murder mycelium.

I can't remember where I got the info on UV-C destroying agar components. :facepalm:

I have done a lot of testing of UV-C on my own and my best conclusion is that it doesn't really do much for mycology.

*I am trying to add to the discussion. I am not trying to cut anyone's ideas down or be the last word on anything.


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OfflineCelestialexplorer1
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Re: Uv-c to sterilize agar [Re: mycorry]
    #26645819 - 05/03/20 11:54 AM (3 years, 9 months ago)

UV Light Sterilizer E-209 - Ultra Violet Sterilizing Cabinet https://www.amazon.com/dp/B0863HK6VY/ref=cm_sw_r_cp_api_i_YtWREbNBACF9S

Here is basically what I’m talking about ghetto rigging up a solid tub or making a trunk maybe. Depends on if I start doing a lot more agar work.


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InvisibleAndyHinton


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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26646492 - 05/03/20 04:52 PM (3 years, 9 months ago)

I wouldn't sterilize media with it. You need to cook thoroughly because encased contaminants on the bottom can easily travel up the sides. It's like drinking from a glass with a dirty stem, bad technique.

You can effectively sterilize a working space such as a polypropylene or polyethylene tub. It doesn't reach shadows. So remember to disinfect everything before loading it in the tub, then run the light.

Here's an example of how to use UV light into your setup. The bulb is a 15W Philips Holland fluorescent-type recycled from a flow hood. Invest in a good non-ozone bulb that you can trust emits 280 nm light.



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Edited by AndyHinton (05/03/20 04:54 PM)


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Invisibleninja cat 09
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Re: Uv-c to sterilize agar [Re: AndyHinton]
    #26647677 - 05/04/20 08:00 AM (3 years, 9 months ago)

Quote:

AndyHinton said:
I wouldn't sterilize media with it. You need to cook thoroughly because encased contaminants on the bottom can easily travel up the sides. It's like drinking from a glass with a dirty stem, bad technique.

You can effectively sterilize a working space such as a polypropylene or polyethylene tub. It doesn't reach shadows. So remember to disinfect everything before loading it in the tub, then run the light.






If you're disinfecting everything, what's the point? Has it helped reduce your contam rate?


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OfflineCelestialexplorer1
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Re: Uv-c to sterilize agar [Re: mycorry]
    #26647939 - 05/04/20 10:01 AM (3 years, 9 months ago)

That sort of input is exactly what I was looking for mycorry thank you. I didn’t know if it would work that’s why I asked but your point makes a lot of sense and it’s something I had thought about also. It seems like if it kills the bad bacteria it could also genetically alter the stuff you want as well. I’ve heard Uvc can alter human dna.


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Edited by Celestialexplorer1 (05/04/20 10:03 AM)


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Offlineleschampignons
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Re: Uv-c to sterilize agar [Re: Celestialexplorer1]
    #26648765 - 05/04/20 05:46 PM (3 years, 9 months ago)



an interesting vid on the subject.

most important of all keep UVC lights away from your eyes and skin. these lights can do permanent damage to you in a short amount of time.


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Invisiblepanne cyanne
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Re: Uv-c to sterilize agar [Re: leschampignons]
    #26650112 - 05/05/20 08:52 AM (3 years, 9 months ago)

i take the dimmest view on UV in grows.
long term eye damage is possible.


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InvisibleAndyHinton


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Re: Uv-c to sterilize agar [Re: ninja cat 09]
    #26650246 - 05/05/20 10:05 AM (3 years, 9 months ago)

Quote:

ninja cat 09 said:
If you're disinfecting everything, what's the point? Has it helped reduce your contam rate?



It's all about keeping track of the relative sterility of different overlapping zones, and modifying your workspace and technique to reduce contamination potential.

Say you have a pressure-sterilized petri dish verified by autoclave tape. This encloses a small sterile area for an instance of microbe culture. It's upstream of things already inoculated, themselves upstream wild specimens, etc. To help ensure sterility, you need to disinfect the outside of it and create a reasonably sterile area to handle it in.

That's where the UV light comes in: assuming a clean and disinfected workspace, it will fry and disintegrate any unshadowed DNA in the workspace, killing all exposed contaminants. PP/PET and similar plastics effectively absorb UV spectra, which is why Plexiglass windows turn opaque in the sun. So the rays are well contained in the workspace.

Before I had to assume a 19th-century standard of sterility and adjust my technique to use syringes and ported lids. Now I can assume a mid-century standard and use regular unmodified stuff handled in sterile air. Same low contamination level, almost always traceable to specific mistakes in technique.

An example of technique failure. Some test cryotubes contaminated because I tried to spin down and combine pellets from 25 mL broth in 1.5 mL tubes (bad idea to clumsily manhandle it with toothpicks). Now I'll do what I've always done, but better: pipette 5 mL ice cold saline, shake vigorously, and resuspend those pellets in cryo medium.


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Edited by AndyHinton (05/05/20 10:08 AM)


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Invisiblepanne cyanne
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Re: Uv-c to sterilize agar [Re: AndyHinton]
    #26650373 - 05/05/20 10:53 AM (3 years, 9 months ago)



unshielded 280nm in an open lab.
your far braver than i.

you should read about blue light UV hazards.
i was taught anything below 320 nm is risky.
PP isnt enough of a shield IMO.

its on the edge of human light perception.
that thing is many time brighter than it appears.
im not sure id be in the room with that even with UV glasses.


Edited by panne cyanne (05/05/20 11:02 AM)


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Re: Uv-c to sterilize agar [Re: panne cyanne]
    #26651015 - 05/05/20 03:04 PM (3 years, 9 months ago)

I work with UV bulbs that are over 6000 watts and produce ozone
I would never consider using UV for at home disinfection of anything.


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Offlinebrikogen
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Re: Uv-c to sterilize agar [Re: bodhisatta]
    #26652839 - 05/06/20 12:11 PM (3 years, 9 months ago)

I don't get the hysteria. With everything care and proper education is important. Just like alcohol can set your house on fire if you are not properly informed on how to handle it and its dangers. SAB explosions, PC accidents, Agar bottle explosions...have happened before due to improper use and careless behaviour. If you are not comfortable with using something, don't.

However as pointed out before agar can be sterilized with the proper UV-C light such as the philips bulbs which are made to not generate ozone. Also make sure you cover it with a light blocking container while it's on. It only take a couple of minutes to sterilize agar plates with UV-C opposed to an hour or more to get your autoclave setup ready, reach pressure, autoclave and cool off. Nevermind the huge energy difference.

At Least give people proper instructions so they don't fuck up. Unless someone is asking about gamma radiation sterilization, then you can tell them to back the fuck away.


Edited by brikogen (05/06/20 12:15 PM)


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InvisibleAndyHinton


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Re: Uv-c to sterilize agar [Re: brikogen]
    #26652924 - 05/06/20 01:07 PM (3 years, 9 months ago)

For the record, you should only use pressurized steam to sterilize media. Each plate is a sacred garden that you must protect from contamination.

Making a sterile environment is essential. A flow hood is probably the best way. The next best way, I think, is to irradiate an enclosed space.

There's no special instructions beyond wiring a lightbulb, using the thickest PP/PET possible, closing the sash while the bulb is lit, and avoiding ozone bulbs.


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Edited by AndyHinton (05/06/20 01:37 PM)


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