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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
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Moldy basement SAB work (now with video)
#27000953 - 10/24/20 08:32 AM (3 years, 3 months ago) |
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Sucks!
Any tips for making this easier? Here is a pic of my setup:

And looking at this pic closely and critiquing my damn self I just realized there is a giant gaping chasm in the middle of the table that is probably causing a giant air current.....
But aside from that massive flaw that I'm going to rectify right now (which may be the root of all my problems) What precautions would you take when working in a moldy basement? Is it okay to be working right off the table? I clean the surfaces in and around the SAB whenever I work, wear gloves to avoid skin flakeage, iso my gloves a couple times while working, flame the scalpel outside the box, keep my deli containers upside down when not working and sideways while working.
Here are the no pours I'm using:

They have a tiny ass hole on the side covered in 3 layers of micropore.
While I'm here, I have a second question. I have had some unopened containers contam on me as well. Not all, but enough to raise an eyebrow. Is this because of my moldy basement? I pour the agar, PC for 45 - 60 mins, let the PC depressurize on its own. Thinking about adding another layer or 2 of micropore. Recipe is 10g agar, 10g LME, 500ml water.
Thanks all
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. “It would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.” -Albert Einstein
Edited by McDominator (10/25/20 10:06 AM)
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
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Is there another area you can work in that isnt so moldy?
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 3 months
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Re: Moldy basement SAB work [Re: eatyualive]
#27001075 - 10/24/20 10:27 AM (3 years, 3 months ago) |
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I wish I could say yes, but I don't have a lot of space and share that space with 2 monster dogs, a kid, and my wife.
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. “It would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.” -Albert Einstein
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 3 months
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It's not the best video, I screwed up a couple times, and I know that plate is fubar. And the damn water on the agar. I don't have high hopes for these plates! I just wanted to give an idea of what my process is to see if you guys think it looks okay or if there are any changes I should make. Thanks!
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. “It would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.” -Albert Einstein
Edited by McDominator (10/25/20 09:23 AM)
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MLPismyOPSEC
That One Ponyfucker


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You know you dun goofed on the second transfer @ 3:06. Other than that, i don't notice anything major. I feel you move your cups too quick, that's most likely where contams are coming in. Moving too fast stirs up the air. The moldy basement doesn't have much to do with contams, because the SAB works very well to keep that shit out.
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 3 months
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Oooh. Moving the cups too quick eh? That's an easy thing to fix. In my mindset of trying to keep things open for less time I'm trying to be quick about things. Guess that didn't translate well to the whole process 😅 Relieved to hear my basement may not be a problem after all.
Yea, and that second plate was being a dick. 
Thanks!
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. “It would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.” -Albert Einstein
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MLPismyOPSEC
That One Ponyfucker


Registered: 11/13/18
Posts: 884
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Fuck that second plate! I always think of the front of the SAB as a big curtain. If there's no motion in the air to disturb the curtain, the air in the SAB stays perfectly* still, and everything falls straight down. If there's nothing above your plates, there's nothing to fall onto them. It is good thinking to not have your plates open long, but time isn't an issue. I leave them sit open in the SAB for minutes at a time.
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IdiotCircusBoy
Human Person



Registered: 09/22/20
Posts: 294
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Is the background noise a washing machine cycling? It almost sounds like a HVAC fan kicks on at like 1:33. Can you run a dehumidifier for a day or so and spray down everything in the basement that looks suspicious with mold and mildew cleaner.
-------------------- Just call me Idiot "People hasten to judge in order not to be judged themselves."
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diegosf
scientific dreamer



Registered: 06/30/20
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Re: Moldy basement SAB work [Re: McDominator] 1
#27003433 - 10/25/20 07:15 PM (3 years, 3 months ago) |
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Hi! Nice to see your work on video.
I also have the same problem of you, with the little pool of water remaining on the plates, so i used to do what you did, let it turned down for a while and then let it drop on the floor of the SAB. But i started to notice a lot of contams poking on the edges of the agar, days after i open them in the SAB and did the transfers..
So i figure it out that it should be probably because of the water remaining on the very little space between the lid and the cup, that was bringing contams inside, through that line of water.. I tested when i keep the plates down, dont turning up again, and they remain clean. My plates are kinda different than your though, there is actually a space where the lid is curved enough to hold contams in there.. Probably when i let the water touch the lid, it gathers the bacteria and spores of mold, and when i close and turn right again, they fall through the sides..
Dont know if its your case, but in the video there is a time when you let some drops of water in the lid, and automatically i thought about what used to happen to me.
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verum subsequentis
seeker of truth



Registered: 03/22/16
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Re: Moldy basement SAB work [Re: diegosf]
#27003725 - 10/25/20 09:40 PM (3 years, 3 months ago) |
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I would recommend Moving a tad more slowly and put a rack of some sort in there to do your work on. The floor of the sab is where all the nasties land. Flipping those bitches over and setting them on the filthy ass floor with the lids cracked and sliding them around is just begging for it. If you don't want a rack, at least put one dish on top of another while working with it.
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A.k.a
Stranger



Registered: 10/27/19
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Yeah I’m with verum. Having the semi open lids on the ground sliding around is probably the problem.
I’d bet either keeping them right side up or the way you did them but keeping them on top of another container would help a lot.
When you have them upside down and press on the bottom to slide them back it seems like the pressure from your fingers would push air out and then suck in all kinds of junk stirred up from moving them when you took your fingers off.
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LAGM2020     
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ModularMind
M.P.F.



Registered: 02/09/10
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Re: Moldy basement SAB work [Re: McDominator] 1
#27003840 - 10/25/20 11:15 PM (3 years, 3 months ago) |
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You can put a wet towel under some cookie cooling racks to trap the falling contaminate spores during your still period. They won’t be wafted airborne by slight hand movements as they would from a solid non-porous surface.
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verum subsequentis
seeker of truth



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Re: Moldy basement SAB work [Re: ModularMind] 2
#27003853 - 10/25/20 11:27 PM (3 years, 3 months ago) |
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Jesus. I don't miss sabs at all. Totally forgot about the ol wet towel.
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ModularMind
M.P.F.



Registered: 02/09/10
Posts: 7,902
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Always feels counterintuitive and non-conducive to sterile procedure but yeah.
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A.k.a
Stranger



Registered: 10/27/19
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I just had to move my SAB before reading this, here’s my set up that works fine.

The rack is crucial. I still keep a couple empty plates stacked on it that I put the ones I’m working on up on too.
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LAGM2020     
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mushboy
modboy



Registered: 04/24/05
Posts: 32,397
Loc: where?
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Re: Moldy basement SAB work [Re: A.k.a] 1
#27004099 - 10/26/20 07:14 AM (3 years, 3 months ago) |
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Quote:
verum subsequentis said: Jesus. I don't miss sabs at all. Totally forgot about the ol wet towel.
i always worked on a hard surface towels are obnoxious af

Quote:
A.k.a said:

eww redbull
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McDominator



Registered: 08/29/19
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Loc: California
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Re: Moldy basement SAB work [Re: mushboy]
#27004116 - 10/26/20 07:29 AM (3 years, 3 months ago) |
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Awesome, thank you guys. Another simple thing to implement! I have a rack I just found that I never use for anything else.
I have about a 70% contam rate right now, but everything you are saying sounds pretty logical as to why that may be the case! 
And I did think about buying or building a flow hood... that might be something I go for in the future.
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. “It would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.” -Albert Einstein
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mushboy
modboy



Registered: 04/24/05
Posts: 32,397
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Re: Moldy basement SAB work [Re: McDominator] 1
#27004119 - 10/26/20 07:34 AM (3 years, 3 months ago) |
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i dont know if any one mentioned it. too lazy to check but i wouldnt recommend using the lid as the floor either.
think the game operation. every time you make contact with the box it vibrates. vibrating box is how shit falls into your work. dont touch the box. the lid is attached to the box. loose the lid.
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Camera93
We got dicks like Jesus



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Re: Moldy basement SAB work [Re: mushboy]
#27004124 - 10/26/20 07:40 AM (3 years, 3 months ago) |
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idk about a wet towel, but I use a towel as a way to kinda brace the SAB and I like feeling like there is a good "seal" around the bottom
-------------------- All I need are some tasty waves, a cool buzz, and I’m fine. Whatever you decide won’t really impact our survival Close your eyes, and do the best that you can
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redbee
prole spore threat


Registered: 03/30/20
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Re: Moldy basement SAB work [Re: diegosf] 1
#27004174 - 10/26/20 08:44 AM (3 years, 3 months ago) |
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Quote:
diegosf said: Probably when i let the water touch the lid, it gathers the bacteria and spores of mold, and when i close and turn right again, they fall through the sides..
Quoted for truth. I'm using similar containers as you, McDominator—same style, but mine are slightly smaller. I've been struggling with contams, but improving. I first tried using these upside-down like pastyplates, and like you do in your video. But I found that I pulled in contams through the condensation water touching the lid and then falling back onto the agar, like diegosf described.
I've switched to using these more like petri dishes, and my contam rate has gone down. I don't EVER flip them upside-down. I try to avoid lifting them off the surface of my rack when I'm transferring. I'll prep a stack of transfers, and crack the lids on each before I flame my tools, then treat them like petri dishes—I lift the lids, make the transfers, set the lids down gently, then re-seal them all once I've finished with a set of transfers. I find that this method also involves way less movement, and I feel like I'm able to keep from stirring the air too much by just lifting lids as opposed to flipping whole plates.
The only time I'll flip them upside-down is if the surface is absolutely covered in water. I'll empty those out quickly, then flip them back right-side up, and do my transfers that way. I then watch those particular plates like a hawk for any contam, for the next few days.
Anyway, thanks a bunch for posting these videos, this has been hugely helpful to see the collective feedback! Keep us posted about how things go, I'm curious to hear what methods you wind up adopting.
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redbee's LAGM 2021
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