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Celestialexplorer1



Registered: 03/25/20
Posts: 1,803
Loc: Floating in a tin can
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Re: Moldy basement SAB work [Re: redbee]
#27004372 - 10/26/20 10:43 AM (3 years, 3 months ago) |
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I don’t know if anyone’s mentioned this but I used to work for/run crew on a mold/flooded rehabilation crew for homes and we would go and spray. Bleach and alchohol only harm the mold they will make it look like it went away but it will recover with a vengeance. They sell microban online in gallons as medic lean and you can get the gallon and a pump sprayer and dilute it with water and spray the whole basement and that WILL kill any mold/spores on the all the surfaces. Just the surfaces but it’s better than nothing. Periodically I’ll take the pump scraper and just spray surfaces in my grow room for updated cleaning.
Just a thought. Sterile tek is key though. You can use a sab in dirty room and still be alright but it’s good to keep it as clean as you can to limit the accidental mishap
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 To spend just one moment in eternity
Edited by Celestialexplorer1 (10/26/20 10:44 AM)
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Celestialexplorer1



Registered: 03/25/20
Posts: 1,803
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Re: Moldy basement SAB work [Re: redbee]
#27004374 - 10/26/20 10:44 AM (3 years, 3 months ago) |
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I don’t know if anyone’s mentioned this but I used to work for/run crew on a mold/flooded rehabilation crew for homes and we would go and spray. Bleach and alchohol only harm the mold they will make it look like it went away but it will recover with a vengeance. They sell microban online in gallons as medic lean and you can get the gallon and a pump sprayer and dilute it with water and spray the whole basement and that WILL kill any mold/spores on the all the surfaces. Just the surfaces but it’s better than nothing. Periodically I’ll take the pump scraper and just spray surfaces in my grow room for updated cleaning.
Just a thought. Sterile tek is key though. You can use a sab in dirty room and still be alright but it’s good to keep it as clean as you can to limit the accidental mishap
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 To spend just one moment in eternity
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
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Re: Moldy basement SAB work [Re: redbee]
#27019063 - 11/03/20 09:57 AM (3 years, 2 months ago) |
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Quote:
redbee said:
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. 
So I made myself a bigger SAB, in preparation for eventual G2G transfers and got myself a rack. Recently did transfers without tipping them upside down. Definitely moved more slowly, but I think my hand went over a dish at one point which is an issue I didn't have with them upside down, need to practice. The other new issue I ran into was where to put the damn lids while working and worrying about getting contams on the lids. Always something. Upside down makes that part much easier, but apparently also contam prone. I made a video of my first time trying this new method and decided not to post it because I have a lot of work to do on the technique that I can already see.
I made 2 plates with a culture syringe (king oyster) and one of them has a few spots of mold while the other is squeaky clean. A syringe drop is much easier than cutting and transferring a wedge and I still botched one So I made 4 transfers out of that dish and we will have to see how those turn out in time.
-------------------- 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, 2 months
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Small update for all who are interested. I made 6 transfers, kept all dishes upside down on raised racks (like 5 inches high). 6 out of 7 dishes became contaminated. (including the source). So now I've stopped going upside down and we will see what happens! Work in progress!
-------------------- 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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Professor X
School for the Gifted



Registered: 04/18/19
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Re: Moldy basement SAB work [Re: McDominator] 1
#27060103 - 11/27/20 12:23 PM (3 years, 2 months ago) |
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I do all my transfers in front of a flow hood 6 feet from a rat cage with 95+% success rate. It's all about technique. Slow deliberate movements, careful positioning and lots of 70% iso. SABs work awesome in bad conditions. Used one in a carpet room for years. Practice makes perfect.
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 2 months
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Finally a good update!
I made 3 transfers to the deli cups, all 3 failed on me. I have close to a 100% failure rate with those after doing close to 100 transfers.
A few days later I made 3 transfers to the Ziploc twist tops..... No contams. Cleanest growth I've had yet. 3 for 3 going strong.
More experimenting needs to be done, but so far the deli containers are going in the trash. My guess is it has something to do with how the covers come off and snap on. Too much movement happening because they get stuck. With the twist caps, there's no snapping or popping going on whenever I maneuver the lids.

Finally feeling a bit hopeful here after months!
-------------------- 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 (12/11/20 06:17 AM)
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 2 months
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Guys, seriously. Went from about a 90% failure rate to a 90% success rate by switching containers. Just wanted to inform you all of what worked in case anyone comes across this. It's probably some sort of user error that was corrected by changing containers. Maybe I don't maneuver the lids of the deli containers as well as others. But this worked, and I'm so stoked. Never felt confident with agar before, but I feel like I'm finally getting there!
-------------------- 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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verum subsequentis
seeker of truth



Registered: 03/22/16
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Congrats
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TheBoJim
Strangest


Registered: 07/23/20
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Quote:
McDominator said: Guys, seriously. Went from about a 90% failure rate to a 90% success rate by switching containers. Just wanted to inform you all of what worked in case anyone comes across this. It's probably some sort of user error that was corrected by changing containers. Maybe I don't maneuver the lids of the deli containers as well as others. But this worked, and I'm so stoked. Never felt confident with agar before, but I feel like I'm finally getting there!
good to hear! and you learned some better techniques along the way. why not go for disposable plates?
Edited by TheBoJim (12/17/20 08:50 PM)
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nektar61
Into SporePlay



Registered: 07/04/20
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Re: Moldy basement SAB work (now with video) [Re: McDominator]
#27194681 - 02/09/21 01:50 AM (2 years, 11 months ago) |
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What about making a little room inside the basement...using a wood frame and plastic of some kind.
They did this on Breaking Bad for that mobile army field hospital that Gus had set up in Mexico. Then Walt did the same thing when they were cooking inside the houses that were tented for bug spraying.
Just make one that's much smaller. Enough for just your chair and a small desk, big enough for your SAB and no more.
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DigDug
Cultivator


Registered: 05/05/20
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Re: Moldy basement SAB work [Re: nektar61]
#27194784 - 02/09/21 04:53 AM (2 years, 11 months ago) |
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Your aseptic technique looks pretty good. I agree with the others. Either do your work on a rack, or soak a paper towel in isopropyl and do your work on there. You'll cut down contams a lot this way. Nice job.
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nektar61
Into SporePlay



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Re: Moldy basement SAB work (now with video) [Re: McDominator]
#27195131 - 02/09/21 09:37 AM (2 years, 11 months ago) |
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Quote:
McDominator said: Sucks!
Any tips for making this easier? Here is a pic of my setup:

I'd tape up the big crack above your right arm hole. at least on the outside, probably with duct tape. Or clear packing tape.
What are the wires and rods on top of your SAB around the weight?
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ModularMind
M.P.F.



Registered: 02/09/10
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Re: Moldy basement SAB work (now with video) [Re: nektar61] 1
#27195424 - 02/09/21 12:10 PM (2 years, 11 months ago) |
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Lightning
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nektar61
Into SporePlay



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Re: Moldy basement SAB work (now with video) [Re: ModularMind]
#27195992 - 02/09/21 05:46 PM (2 years, 11 months ago) |
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Quote:
ModularMind said: Lightning
Ah. I'll try that. Duh. Why did I never think of that. That would help a lot. Thank you.
I thought it was something electrostatic, that would suck any airborne particles to the inside top of the SAB.
Wonder if that would work? It probably pull your spores up there too, and maybe make a tiny unwanted breeze.
I know you've been around here more than a decade longer than me, but I have a thought: I wonder if you can you make this work with a smaller weight on top? I am guessing that is contributing to the crack above the right hand hole, or will eventually.
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ModularMind
M.P.F.



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Re: Moldy basement SAB work (now with video) [Re: nektar61]
#27196828 - 02/10/21 06:26 AM (2 years, 11 months ago) |
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Now that you’ve quoted it, I realize I typo’d lighting. I presume the weight is an effort to keep the box still if bumped. Not sure.
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Professor X
School for the Gifted



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Re: Moldy basement SAB work [Re: ModularMind]
#27197086 - 02/10/21 10:15 AM (2 years, 11 months ago) |
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I see two things, use a wire rack to Jack your plates up off the floor of the SAB, I use a small dish rack with the legs cut off. The other thing I see is you are turning over the recipient plates too much. Don't worry about the donor plate but keep the recipient upside down the entire time. Any plates I turned over like yours I would set to the side and Mark when I used a SAB, most of them failed. One last thing, I watched you sanitize then immediately put your hands over the plates, not good.
I would also make some "sleeves" I use dishwashing glove cuffs cut away from the hands, 2 sets to cover my forearms and iso everytime you move out of the box. Get a torch that stays lit too.
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