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PutACapInHisAss
Stranger Than Fiction



Registered: 08/11/07
Posts: 252
Loc: Earth
Last seen: 6 years, 4 months
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: Pastywhyte]
#23438090 - 07/13/16 03:15 PM (7 years, 6 months ago) |
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blindingleaf said: 10-20mminutes??? no way, I got shit to do man!! like swiping left . I think flaming is easier dude.
My idea isn't to wait 10-20 minutes like in the case of surface sterilization mentioned in this article.
My idea is to start with flame sterilized tools and then keep them in the SAB using the solution in a convenient way between each action. The tools must be set down at times for opening containers, wiping down containers, etc. The tools can be set in a shallow container of this solution or possibly wrapped in a towel or cloth that is well soaked with it.
I can understand your reluctance. All I can tell you is that I have had significant success in the past without flame sterilization of syringes between procedures. Long ago I didn't realize that it was something meant to be repeated between each jar. I just did it once at the beginning. I still had a 90%+ success rate on jar colonization. Heck, as already mentioned, some people claim significant success in open air! It is not that outlandish of an idea, especially in a SAB or using a LFH.
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Mad Season said: So wait they're only focusing on pathogens? What about the spores that we have worry about that can EASILY survive well any sanitizer. Also the resource says virtually every thing. Not everything (which flame guarantees).
First, the word pathogen is my word, it is not actually in the article. Second, in their trial they claim “the acidified solution killed ALL of the spores on all of the surfaces.” “All” is not “virtually all,” even though it is said in a later quote. Perhaps the use of the phrase “virtually all" is because in that sentence they say 10-20 mins, rather than the 20 mins where all were killed. I am talking about using it on a surface that should be extremely clean in the first place. Finally, “spores” means spores “that we have worry about.”
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Mycologist217 said: I mean I can buy denatured alcohol clean burning fuel for so cheap that it wouldn't even be economical to use anything else to sterilize my stuff
Maybe it is because of the economy where you live. In the US bleach and vinegar are extremely cheap by the gallon—about $2 for a gallon of vinegar and $3 for a gallon of bleach. A quart of denatured alcohol is $8-10.
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Pastywhyte said: If the chemicals won't hurt the fungi, they won't do the job. We have more to worry about than just spores and endospores. Mycelium fragments from molds are everywhere and if the chemical won't kill those, that it is ineffective. If it can kill them then it will harm your culture as well. People have been dipping biopsy tissues in all sorts of things for ages in an attempt to clean them before putting to media. The result is always damage to the intended culture.
This topic comes up at least twice a year. So far I still remain unconvinced. If you still want to try then go for it. But understand why it's considered bad practice.
This is a very good point, I hear you—fragments of mold (etc) would still be a problem, no doubt!
However, it is a small thing to wipe off the end of a scalpel with a sanitary towel after each procedure. I do this already and it is very likely to remove most or all such fragments.
Still, I really think it is a question of what is good enough. I am not trying to save people’s lives using tissue cultures. Also, I along with many others have had significant success with far less than optimal procedures. If it works consistently with a high rate of success then there is no argument.
Plus, while you may have an aversion to such relatively innocuous chemicals, I have an aversion to fire and moving my hands in and out of my SAB. I think the air and my body (which is also outside of the SAB) are as likely vectors for contamination as my tools.
And fire with alcohol in the SAB is a certain danger. Although, with such a possibly effective non-flammable sanitizer, I can think of no reason to continue using isopropyl in the SAB, or for wiping anything down, especially when it is also much more expensive.
I may feel differently when I am using a LFH in the future. If the air is clean and my gloves become less likely a vector because they are kept behind the containers in the airflow, what does it matter if they are contaminated while flaming outside of the clean space.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: PutACapInHisAss]
#23438141 - 07/13/16 03:43 PM (7 years, 6 months ago) |
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Quote:
PutACapInHisAss said:
Quote:
Mad Season said: So wait they're only focusing on pathogens? What about the spores that we have worry about that can EASILY survive well any sanitizer. Also the resource says virtually every thing. Not everything (which flame guarantees).
First, the word pathogen is my word, it is not actually in the article. Second, in their trial they claim “the acidified solution killed ALL of the spores on all of the surfaces.” “All” is not “virtually all,” even though it is said in a later quote. Perhaps the use of the phrase “virtually all" is because in that sentence they say 10-20 mins, rather than the 20 mins where all were killed. I am talking about using it on a surface that should be extremely clean in the first place. Finally, “spores” means spores “that we have worry about.”
No it doesn't, it means dried bacterial spores, and it doesn't even list what types of them. I'm talking broad scale of everything. Saying it killed every bacterial spore they put on a surface doesn't mean much to me. Tell me what it killed and how long it took. Furthermore it takes 20 minutes to kill everything. So basically when we wipe it every minute, it'll just add the spores that are on it to the blade. There's a reason why they call it a sanitizer (disinfectant) over a sterilizer. Flame instantly kills everything.
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Miner and his colleagues compared the ability of alkaline (pH 11) and acidified (pH 6) bleach dilutions to disinfect surfaces contaminated with dried bacterial spores, considered the most resistant to disinfectants of all microbes. The alkaline dilution was practically ineffective, killing all of the spores on only 2.5 percent of the surfaces after 20 minutes. During the same time period the acidified solution killed all of the spores on all of the surfaces.
"Diluted bleach at an alkaline pH is a relatively poor disinfectant, but acidified diluted bleach will virtually kill anything in 10 to 20 minutes," says Miner. "In the event of an emergency involving Bacillus anthracis spores contaminating such environmental surfaces as counter tops, desk and table tops, and floors, for example, virtually every household has a sporicidal sterilant available in the form of diluted, acidified bleach."
My point being that there's much more to this than worrying about bacteria spores, which is what they focused on. And after 20 minutes they'll still be around, you should have gone through how many plates by then?
Edited by Mad Season (07/13/16 04:10 PM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: PutACapInHisAss]
#23438159 - 07/13/16 03:51 PM (7 years, 6 months ago) |
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You can use soap and water to wet the SAB 
End of the day it's your grow. I wish you well.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: Pastywhyte]
#23438212 - 07/13/16 04:12 PM (7 years, 6 months ago) |
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peroxyacetic acid + 26% h2o2 sanitizer basically will sterilize stainless steel surfaces if they're surgically clean before hand IE no soil at all, and the stainless is passivated and ready to go. but it's still a sanitizer.
diluting bleach in water acidifies it. 1 part bleach to 9 parts water is a very effective sanitizer too nearly a sterilizer on already surgically clean hard surfaces
doesn't really matter though with a SAB or even a FH to use a bunch of sanitizer or one that's particularly better than another. 70% iso works fine for wiping your jars down.
the biggest problem you'll have is eliminating yourself as a contaminant vector and you can't sterilize yourself even with gloves and masks or showering before hand. so to worry about the chemicals you're using totally misses the point, if you're having problems fix yourself not your chemical regimen. because the sanitizers are not saving your ass.
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Ginge
Sponge Brain


Registered: 07/08/18
Posts: 202
Loc: Rocky Mountains
Last seen: 5 years, 9 days
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: bodhisatta]
#25381087 - 08/11/18 07:38 AM (5 years, 5 months ago) |
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Okay, first of all. Your counter arguments are respectful and thought out, from your perspective. However, you asked for the perspective of other growers. While ingenious new things are being discovered every day, millions of supposedly ingenious new things are failing. I love experimentation, and I love taking what I have learned about mycology and applying it to the variety of situations I encounter. BUUUUT, I have had a lower success rate with my own ideas or combinations of shroomery teks that I thought worked together, than I have had just following teks exactly. Flaming your loop, needle or blade is THE transfer tool tek, if you come up with something more effective, CHEERS, but it has cost me a lot of time, money and fruits to experiment. I also really think there is a lot of value in what Bod says about fixing the contamination vectors using known techniques instead of recreating something to compensate for those vectors. You would be in a much better realm to experiment if you had really good technique to add to your new ideas. I applaud your early success with plates, I had 1 contam of 32 plates on my first MS to agar run, 8 did not germinate however. I will be working with syringes tonight on agar, and I am hoping this is the last time I work with a vendor syringe! (unless I pc them for my LC's )
-------------------- Ginge
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Agar Plates...Am I Lucky Or...? How is Your Plate Contam Rate? [Re: Ginge]
#25381098 - 08/11/18 07:46 AM (5 years, 5 months ago) |
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Check thread ages. You're asking questions to people who haven't been around in years and probably ain't coming back. The other thread you bumped was 7+ years old for an easy agar question.
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