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 CarmeloLabadie
Stranger
Registered: 09/13/21
Posts: 1
Last seen: 1 month, 5 days
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 No pour agar sterilization methods 
#27466296 - 09/13/21 02:40 AM (1 month, 5 days ago) |
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I'm wanting to get into agar work but the first two attempts didn't go very well. I watched loads of videos and did the sterilize and pour method first. Even though I cleaned the SAB, gloves and sleeves I still got about 70% contaminated. So I tried the no pour method next. After taking everything out of the PC I had loads of excess water in everything even after carefully wrapping with foil. I also had what appeared to be algae in the agar but attribute that to letting the agar boil over on the stir plate, that I can fix.
I'm using a mixture of 60 and 90 mm glass petri dishes for doing culture samples and condiment cups for making agar to grain transfers. I have some jars I can put the 60 mm dishes in but the 90's are too big for them or most other jars for that matter. On a side note the 90's stack better than the 60's and that makes a difference when pouring the agar. I'm pushing it at 5 tall with the 60's before they want to topple. I've looked into the Ziploc twist n lock containers but the 60's want to tip over and the 90's are to big.
What methods do you use to keep moisture infiltration to a minimum during PC'ing?
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Carmodycaramel
Caramelized
Registered: 04/03/17
Posts: 1,179
Loc: United States 
Last seen: 3 hours, 48 minutes
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Hard for me to understand but what do your no pour containers look like exactly? Are you placing pre poured petri dishes inside of jars and pressure cooking?
EDIT : Maybe I'm just a dunce.
My method for no pour involves 4oz mason jars with a single hole poked in the lid covered in micropore tape. I prepare the agar in the microwave, pour into the jars, screw the lids on and then cover the lids in tinfoil and pressure cook.
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/0---0---0---0---0---0---0---0---0---0\
\0---0---0---0---0---0---0---0---0---0/
Edited by Carmodycaramel (09/13/21 02:57 AM)
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CreonAntigone
Stranger
Registered: 05/31/21
Posts: 456
Last seen: 5 hours, 52 minutes
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It's not entirely clear - what kind of container are you doing the no-pours in? Are you just PCing the glass Petri dishes? For how long?
If algae germinates on your plates then they aren't sterile. A proper PC cycle will definitely kill the algae spores. Perhaps you aren't sterilizing long enough or perhaps your PC isn't functioning correctly.
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alaskappalachian
peakbagger
Registered: 10/22/19
Posts: 1,355
Loc: The 49th Dimension
Last seen: 1 hour, 1 minute
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Re: No pour agar sterilization methods [Re: CreonAntigone]
#27466319 - 09/13/21 03:09 AM (1 month, 5 days ago) |
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When pouring your agar, let it cool to about 120 before pouring and that will take care of a lot of your problem with moisture. With prepours, prep your agar and pour, then leave those lids off until the dishes cool THEN lid the plates/dishes and PC. I have almost zero condensation in mine without any additional work. Also, sanitizing things with isopropyl is good, but won't in the least bit compensate for improper technique. Not an insult at all; just you are new and need a lot of practice. Go back and read Bohdi's agar teks and some of the prepour teks n the sticky threads. They cover all of this and more. Best of luck. You do a few runs and actually read those teks CAREFULLY and you'll be transferring to your heart's content in no time. You got this.
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"If only you could see what I've seen with your eyes..."
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 DERRAYLD
Constructus
Registered: 05/13/02
Posts: 3,403
Loc: South Africa
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No pours have to be cooled slowly to avoid condensation or you have to wrap the pc or similar to allow the agar to cool slowly.
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alaskappalachian
peakbagger
Registered: 10/22/19
Posts: 1,355
Loc: The 49th Dimension
Last seen: 1 hour, 1 minute
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Re: No pour agar sterilization methods [Re: DERRAYLD]
#27466335 - 09/13/21 03:25 AM (1 month, 5 days ago) |
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Untrue. As I just mentioned, just let the prepours cool before lidding and you're gravy. I keep about 150 at a time in rotation and pull those right out after PCing. Every time.
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"If only you could see what I've seen with your eyes..."
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