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Bertoxulous
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Registered: 04/26/16
Posts: 23
Last seen: 8 years, 8 months
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Help me with my SAB work procedures please
#23202094 - 05/08/16 11:42 AM (8 years, 8 months ago) |
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I've read so much through the years with so many differing of opinions.
I was doing some SAB work over the past couple weeks, and it always seems to boggle me what order I should be doing things in.
Does this seem "proper"?
1. Wipe work surface with alcohol 2. Place towel down on work surface and spray with soap water 3. Place everything I need in the box on the towel after wiping with alcohol. (except for tools and syringes that will be flamed) 4. Spray SAB while inverted with soap water, then place over all my materials. 5. Run my HEPA air purifier for an hour or two, then shut off and wait an hour. 6. Take shower 7. Glove up and wipe hands with alcohol. 8. Begin working.
One other concern I had was regarding flame sterilizing out side of the box. Is it better to flame sterilize everything prior to starting and place in the box, or sterilize as I go? It seems like moving hands in and out of the box a lot to flame things would create a lot of air currents.
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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: Help me with my SAB work procedures please [Re: Bertoxulous]
#23202103 - 05/08/16 11:43 AM (8 years, 8 months ago) |
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5 and 6 should happen first... And just flame as you go... Your scalpel should stay glowing as you enter the SAB..
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Bertoxulous
Stranger

Registered: 04/26/16
Posts: 23
Last seen: 8 years, 8 months
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Re: Help me with my SAB work procedures please [Re: LocN9ne]
#23202108 - 05/08/16 11:46 AM (8 years, 8 months ago) |
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OK, so scrub the air in the room and shower, before even setting anything up?
That makes sense.
While I'm at it, I'm about to inoculate some agar for the first time with some Pasty plates. Do I need to wrap the lids of the glad containers with parafilm/plastic wrap?
Edited by Bertoxulous (05/08/16 11:48 AM)
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Josex
#cheat_code



Registered: 11/13/15
Posts: 8,999
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Re: Help me with my SAB work procedures please [Re: Bertoxulous]
#23202147 - 05/08/16 11:58 AM (8 years, 8 months ago) |
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Wow I love that you have it all correct right from the start 
Personally, I don't wipe my SAB with alcohol. Instead I pour some soapy bleach and with a dish cloth I spread it on the walls of the SAB, being carefull nothing is left dripping.
I think these links might help you a lot if you don't have everything perfectly clear yet.
https://www.shroomery.org/forums/showflat.php/Number/21889950 https://www.shroomery.org/forums/showflat.php/Number/20048771 https://www.shroomery.org/forums/showflat.php/Number/23058476
Edited by Josex (05/08/16 12:13 PM)
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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: Help me with my SAB work procedures please [Re: Bertoxulous]
#23202180 - 05/08/16 12:07 PM (8 years, 8 months ago) |
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Quote:
Bertoxulous said: OK, so scrub the air in the room and shower, before even setting anything up?
That makes sense.
While I'm at it, I'm about to inoculate some agar for the first time with some Pasty plates. Do I need to wrap the lids of the glad containers with parafilm/plastic wrap?
No, because if you made them right they will have a hole covered with micropore tape...
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Q&A
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The more I learn, the less I know.
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Bertoxulous
Stranger

Registered: 04/26/16
Posts: 23
Last seen: 8 years, 8 months
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Re: Help me with my SAB work procedures please [Re: LocN9ne]
#23203565 - 05/08/16 07:42 PM (8 years, 8 months ago) |
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They do have a hole covered with micropore.
My thought was that those containers aren't necessarily an air tight seal, and contams could possibly sneak in?
Maybe I'm mistaken and they are tighter than I think and no need for concern?
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Josex
#cheat_code



Registered: 11/13/15
Posts: 8,999
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Re: Help me with my SAB work procedures please [Re: Bertoxulous]
#23203589 - 05/08/16 07:46 PM (8 years, 8 months ago) |
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Quote:
Bertoxulous said: They do have a hole covered with micropore.
My thought was that those containers aren't necessarily an air tight seal, and contams could possibly sneak in?
Maybe I'm mistaken and they are tighter than I think and no need for concern?
Glad mini rounds? Nope, nothing will get in. Tons of people here are using it without issues. I can't get mini rounds where I live but you can use any type of small pp5 containers with a snap-on lid (or ziplock lid) to do agar, as long as it's fairly high quality.
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Supalemonhaze
Spore syringe hater.



Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: Help me with my SAB work procedures please [Re: Bertoxulous]
#23203611 - 05/08/16 07:51 PM (8 years, 8 months ago) |
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Yeah, they seal quite well enough to keep contams out. Contams can't travel by themselves and they rely on air currents so it's improbable that they will get under a lid and then up to pass between the lid and the bottom. People leave they petri dishes unwrapped when they pour agar until they cool and they have literally no seal without parafilm.
Once you get used to a SAB, some of the steps you do become unimportant. Like the air purifier, wiping box with iso or taking a shower right before you work.
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Munchauzen


Registered: 06/22/11
Posts: 14,348
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Re: Help me with my SAB work procedures please [Re: Bertoxulous]
#23203615 - 05/08/16 07:52 PM (8 years, 8 months ago) |
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Quote:
______Preparing a Still Air Box Workspace______
more of my videos in my signature
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