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fraggy
Stranger



Registered: 02/12/09
Posts: 210
Last seen: 2 years, 4 months
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Noob: Steam sterilizing petri dish with agar inside?
#19239631 - 12/07/13 06:05 AM (10 years, 1 month ago) |
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Sorry if this question has been answered elsewhere, it's just that it's not easy getting search results with "steam sterilizing petri dishes with agar inside"...
I know standard procedure for agar petri dishes is to pour hot liquid agar into sterile petri dishes, under sterile conditions, but I live in a very contaminated place. Forget about buying premade agar with nutrients and even less premade dishes (sterile petri dishes with agar-mix already inside) because they're impossible to get where I live, so I will have to cook up (litteraly) everything myself! Note that these dishes will be for SPORES! So no peroxide allowed here!
I got a glove box, but it is small and uncomfortable, it will do for making spore syringes, and cake inoculations, but I fear it won't be spacious enough for making agar plates, unless I do a very small quantity at a time... Perhaps I should try anyway, but I'd hate to waste material...
So here comes my question:
Would it possible to pour the agar into the petri dishes under non-sterile conditions first, let the agar harden and cool off, place dishe's cover on top, pack the complete dish in aluminium foil and then steam sterilise the whole thing?
Remember this is for spores, so no peroxide allowed...
This method would make my life a lot easier! Would it work?
Anybody ever tried?
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BlackPeace
Self proclaimed CEO



Registered: 11/18/07
Posts: 1,069
Loc: Canada
Last seen: 8 months, 22 days
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Re: Noob: Steam sterilizing petri dish with agar inside? [Re: fraggy]
#19239654 - 12/07/13 06:22 AM (10 years, 1 month ago) |
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The problem is that when the petri cool off, it will suck in dirty air and contaminate. If I were you, I would buy 125ml jars, fill with agar sterilise (with a polystuff filter material on the lid).
The only thing I don't know is how good the texture will be at the end. Everytime I sterilise my agar bottles, I clearly see that most of the agar is in the bottom of the bottle. So I need to swirl it before I pour it.
Maybe try doing that, put a bit more agar in your jar than you would for a petri dish, and swirl it before you leave it to cool.
Good luck, try to search for jar agar making teks, I'm sure you will find a few here.
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BlackPeace
Self proclaimed CEO



Registered: 11/18/07
Posts: 1,069
Loc: Canada
Last seen: 8 months, 22 days
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Re: Noob: Steam sterilizing petri dish with agar inside? [Re: BlackPeace]
#19239674 - 12/07/13 06:30 AM (10 years, 1 month ago) |
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Woah, just made my 1000's post here haha
Feels like, somekind of an achievement or something. I would celebrate but it's way too early for a beer lol
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FooMan



Registered: 02/02/05
Posts: 8,957
Loc: Earth
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Re: Noob: Steam sterilizing petri dish with agar inside? [Re: fraggy]
#19239727 - 12/07/13 07:14 AM (10 years, 1 month ago) |
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I would get a bigger glove/still air box personally. One can be made from a cardboard box lined with plastic like so:
  
Cooking agar on the plates is a pain in the ass. You'd need autoclavable dishes to start, standard plates will melt. The agar may boil over and the condensation inside of the dishes will be horrible. If you need to sterilize inside of the container you may want to look into using PP5 deli containers or short, wide mouth mason jars instead.
--------------------
Quick WBS Prep
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fraggy
Stranger



Registered: 02/12/09
Posts: 210
Last seen: 2 years, 4 months
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Re: Noob: Steam sterilizing petri dish with agar inside? [Re: FooMan]
#19240469 - 12/07/13 11:35 AM (10 years, 1 month ago) |
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Thanks guys, really useful tips, all of them. I happen to have 3 glass petri dishes, the suck in air issue it's a good thing you point it out, that one escaped me. Perhaps that might be solved some way or another, with a towel or some micro pore tape... the condensation would be the biggest issue I suppose. I'm gonna give it a shot to convince myself that it's no good
I did like very much the jar option. Easy to get bye stuff, whatever thing that's transparent and heatproof. I'll try some of those pee-pots that are used for urine samples. Make a pair of holes in the lid, cover holes with micro pore tape and 1cm of agar will do, in order not to waste too much material...
I was thinking of doing some LC to start out with some spores, together with BRF cakes and perhaps do some agar later. But I think agar would be somehow easier for me and give me more control over eventual mistakes I might make, as agar has the benefit of making isolation of myc from contams possible, even if that would mean some more work :P
And the pics of your glove box are greatly appreciated! Cool thing to know a poor man's glove box can be made out of these materials, with a nice artisanal result. Scoring for style points too  I know the idea is to create a "still air" environment, but sometimes I forget, lol!
Still so much to learn!
These things make our hobby so much more enjoyable
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Diamorphine
Constantly contaminated

Registered: 05/01/16
Posts: 121
Loc: Carolina's
Last seen: 6 years, 5 months
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Re: Noob: Steam sterilizing petri dish with agar inside? [Re: fraggy]
#23295468 - 06/01/16 09:32 AM (7 years, 7 months ago) |
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Did this ever work out for you?
-------------------- Try mixing different colored airheads together when tripping it creates new colors and gradients. And they're tasty...
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