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dr0384



Registered: 07/17/10
Posts: 391
Loc: Australia
Last seen: 4 months, 11 days
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Contam problems
#13208070 - 09/17/10 03:36 AM (13 years, 4 months ago) |
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I made an agar transfer. I used antibacterial surface spray in my glove box. then some iso alcohol 91%, I got the antibacterial air spray and covered my bedroom and sprayed some up the arm holes of my glove box. I put tyvek arms on, surgical gloves. Washed my gloves with iso. Washed the zip lock bags with iso then put them in the GB. opened the bags in the GB and took out both petri dishes. Made the transfer. Looked to day the transfer is going like crazy but so is that fucken green mold. WTF. What other precaution can I take? Frustrated
-------------------- Good judgement comes from experience. Experience comes from bad judgement. Be yourself whoever that is because those who care don't matter and those who matter wont care.
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: Contam problems [Re: dr0384]
#13208255 - 09/17/10 06:02 AM (13 years, 4 months ago) |
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You can close the door to the room for several hours before you do the work.
It's more important that the air be still than you spray it with aerosol. I'm very skeptical that you can 'sterilize' the air with an aerosol spray. I think it's more important that the air be still.
also, the inside of a ziplock is sterile. no need to mess with it.
elaborate on the transfer technique? did u flame?
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
Edited by anonjon (09/17/10 06:03 AM)
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LeopardMan
Constantly changing


Registered: 09/21/09
Posts: 5,463
Loc: A tree house
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Re: Contam problems [Re: anonjon]
#13208266 - 09/17/10 06:08 AM (13 years, 4 months ago) |
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Quote:
anonjon said: I'm very skeptical that you can 'sterilize' the air with an aerosol spray. I think it's more important that the air be still.
You can't sterilize anything with an aerosol spray. Sanitize maybe, but certainly not sterilize.
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You have to die a few times before you can really live. -Charles Bukowski-
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andymc
cocoa beetles from zanzibar



Registered: 11/25/09
Posts: 2,395
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Re: Contam problems [Re: dr0384]
#13208304 - 09/17/10 06:30 AM (13 years, 4 months ago) |
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Despite our best efforts, agar dishes still contaminate sometimes. The good news is that you can do another transfer to get your culture out of that contaminated dish, and if you work carefully, the green won't follow it.
All these precautions you've taken are really good, but they'll never have 100% success. Still, they're worth doing -- just imagine how bad contamination rates can be when you don't make efforts to work cleanly.
I find agar work to be the most difficult to keep contamination-free.
Most of us don't have the dough or space for a flow hood
-------------------- How I make spore prints Trade List My flow hood If he asks me "Did you have a good time?" I'll say, "Get the lights, Mr. Grim Reaper" -odds
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NineInchNails
Stranger

Registered: 03/01/09
Posts: 1,190
Last seen: 1 year, 23 days
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Re: Contam problems [Re: andymc]
#13208335 - 09/17/10 06:50 AM (13 years, 4 months ago) |
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I agree with anonjon 100%. You can spray & wipe down everything, but that doesn't do much for what microscopic particles (contam) that is suspended in the air within the GB itself.
Cleaning the GB is not a bad idea, but I only do that 1 out of 4 times that I use mine.
Kill all fans in the room for sure.
put all your stuff inside the GB and let it sit for a good 15-20 minutes. Doing this allows the particles suspended in the air within the GB to settle out.
Flame sterilize and go to work.
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Out Grow
Community Vendor



Registered: 09/03/09
Posts: 917
Loc: midwest
Last seen: 3 months, 26 days
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I'm not saying your breathing out trick or anything but I always where a surgical mask. Figure it cuts down on airborne bacteria and also less disturbing the air.......and of coarse tho obvious one Flow Hood.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Contam problems [Re: Out Grow]
#13208389 - 09/17/10 07:24 AM (13 years, 4 months ago) |
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A flowhood is not a substitute for good sterile technique. If you can't ride a bike, don't get a motorcycle.
-------------------- You make it all possible. Doesn't it feel good?
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dr0384



Registered: 07/17/10
Posts: 391
Loc: Australia
Last seen: 4 months, 11 days
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Re: Contam problems [Re: Doc_T]
#13208435 - 09/17/10 07:50 AM (13 years, 4 months ago) |
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I work in the outskirts of Melbourne. A lot of country work. Therefore I bring home a but load of spores with me. My contamination rate it pretty high. A lot higher than I would like it to be.
I turned the ducted heating off and let settle for about 15 min. I didn't flame as it was a new sterile scalpel. I only partly opened the petri dish so I could access the agar. Cut what I needed and transferred to the agar dish I had next to it. My GB just has holes cut out the side. Doesn't have any kind of seal. Think I might put some kind of seal on it to stop airflow. Any ideas? I also have a little toilet room in my house. I was worried about bacterial contamination from that room, but I'm now thinking if I load all my gear up in there, spray like all hell, have a shower, antibacterial wash, new clothes. It has to be the stillest room in the house and exposed to the least amount of spores.
That will limit it. Is there any way of stopping airborne spores other than h202. I could put some H202 in and just wait the extra time. I have done 20 dishes and 13 have contaminated with green titch or whatever. a couple did bacterial contam. That green shit will be the death of me. Can you h202 the air?
-------------------- Good judgement comes from experience. Experience comes from bad judgement. Be yourself whoever that is because those who care don't matter and those who matter wont care.
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: Contam problems [Re: dr0384]
#13208633 - 09/17/10 09:10 AM (13 years, 4 months ago) |
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You're on the wrong track.
The amount of spores where you are is irrelevent. Wherever there is dirt there is trich. You don't have any more or less than anybody else.
Spraying the air with h202 would be even more futile than the aerosol because the particles are even bigger.
The air doesn't settle in 15-20 minutes. Close the door to the room, close the ducts and windows and let the air settle out for a couple hours. Don't then go back and start spraying stuff.
Don't put an air seal on your still air glove box. You'll disturb the air even worse with that. It'll be an awkward pain in the ass process and you'll have to flame your needle inside the box, which is not recommended.
Don't assume that you 'did everything right' and start looking outside standard technique for corrective measures. Perfect the standard sterile technique. Anything else is just a total waste of your time and resources.
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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dr0384



Registered: 07/17/10
Posts: 391
Loc: Australia
Last seen: 4 months, 11 days
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Re: Contam problems [Re: anonjon]
#13210808 - 09/17/10 06:01 PM (13 years, 4 months ago) |
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kool. thx. So my approach next time will be to move all my equipment that I'm going to use into the toilet. Spray with bacterial aerosol and leave for 2 hours to let it settle, and any spores to settle too. During that time have a shower, get clean clothes on. (buy a face mask) go in do my business and get out. Hope for the best.
-------------------- Good judgement comes from experience. Experience comes from bad judgement. Be yourself whoever that is because those who care don't matter and those who matter wont care.
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LeopardMan
Constantly changing


Registered: 09/21/09
Posts: 5,463
Loc: A tree house
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Re: Contam problems [Re: dr0384]
#13210846 - 09/17/10 06:10 PM (13 years, 4 months ago) |
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Yes but not in the bathroom. Go in the bedroom.
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You have to die a few times before you can really live. -Charles Bukowski-
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dr0384



Registered: 07/17/10
Posts: 391
Loc: Australia
Last seen: 4 months, 11 days
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My bedroom only has 1 door. outside. And I just figure I can bring anything in with me as I come in. The biggest problem in the bathroom is bacteria yea? I thought if I coat every square inch of the air with bacterial aerosol on my way out that should combat the problem. This is what I used to do and never had a contamination. Got lazy in my bedroom and started getting trich. Still think its a bad idea?
-------------------- Good judgement comes from experience. Experience comes from bad judgement. Be yourself whoever that is because those who care don't matter and those who matter wont care.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Contam problems [Re: dr0384]
#13210954 - 09/17/10 06:31 PM (13 years, 4 months ago) |
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Bacteria don't fly through the air by themselves. Like white men, bacteria can't jump.
-------------------- You make it all possible. Doesn't it feel good?
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dr0384



Registered: 07/17/10
Posts: 391
Loc: Australia
Last seen: 4 months, 11 days
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Re: Contam problems [Re: Doc_T]
#13211111 - 09/17/10 07:05 PM (13 years, 4 months ago) |
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Quote:
Doc_T said: Bacteria don't fly through the air by themselves. Like white men, bacteria can't jump.
LOL. So I've done it in my toilet. Not the most bacterial friendly place but I covered the place with antibacterial wash and not going in there for 1.5 hrs. Wait for it to settle. Just before my time is up I will have a shower, put on some clean clothes, put on my gloves and arm guards, make up a mock up face guard, and go in without disturbing the air much and pour my agar. Put the dishes in a zip lock bag and do again tomorrow for my transfer. Maybe later to day if I have time.
-------------------- Good judgement comes from experience. Experience comes from bad judgement. Be yourself whoever that is because those who care don't matter and those who matter wont care.
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: Contam problems [Re: dr0384]
#13211483 - 09/17/10 08:37 PM (13 years, 4 months ago) |
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It seems to me that every time someone says they gonna work in the bathroom, someone goes on about the bacteria. But more often than not it works out fine.
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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LeopardMan
Constantly changing


Registered: 09/21/09
Posts: 5,463
Loc: A tree house
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Re: Contam problems [Re: anonjon]
#13212867 - 09/18/10 04:49 AM (13 years, 4 months ago) |
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Quote:
anonjon said: It seems to me that every time someone says they gonna work in the bathroom, someone goes on about the bacteria. But more often than not it works out fine.
Maybe because in the bathroom there are bacteria, molds and fecal spores? I wouldn't set up my homemade laboratory in the same room where I take a dump
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You have to die a few times before you can really live. -Charles Bukowski-
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Quote:
LeopardMan said:
Quote:
anonjon said: It seems to me that every time someone says they gonna work in the bathroom, someone goes on about the bacteria. But more often than not it works out fine.
Maybe because in the bathroom there are bacteria, molds and fecal spores? I wouldn't set up my homemade laboratory in the same room where I take a dump 
As if your bed, covers, pillows, clothes, and bedroom carpet aren't a giant mold factory?
And fecal spores? When a fecal spore lands, does it germinate into a poop?
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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LeopardMan
Constantly changing


Registered: 09/21/09
Posts: 5,463
Loc: A tree house
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Re: Contam problems [Re: anonjon]
#13212939 - 09/18/10 06:00 AM (13 years, 4 months ago) |
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Quote:
anonjon said:
Quote:
LeopardMan said:
Quote:
anonjon said: It seems to me that every time someone says they gonna work in the bathroom, someone goes on about the bacteria. But more often than not it works out fine.
Maybe because in the bathroom there are bacteria, molds and fecal spores? I wouldn't set up my homemade laboratory in the same room where I take a dump 
As if your bed, covers, pillows, clothes, and bedroom carpet aren't a giant mold factory?
As you probably know molds thrive in warm, damp, and humid conditions and there is a lot of water and humidity in my bathroom. Not in my bedroom Fecal spores are carriers for bacteria I know spores and bacteria are everywhere but some places, like bathrooms and kitchens, have higher concentrations than others. That's a fact
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You have to die a few times before you can really live. -Charles Bukowski-
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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What is a fecal spore?
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Contam problems [Re: anonjon]
#13212963 - 09/18/10 06:22 AM (13 years, 4 months ago) |
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Quote:
anonjon said:
Don't assume that you 'did everything right' and start looking outside standard technique for corrective measures.
This is a big deal.
-------------------- You make it all possible. Doesn't it feel good?
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