How to Pour Agar Plates
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- čas přidán 16. 12. 2009
- Dan gives a demo of how to pour agar plates.
1) Microwave the agar until melted
2) Cool down for 15-20min (until it's cool to touch)
3) Add antibiotics and swirl gently
4) Pour and let sit until solid
5) Store at 4 degrees Celsius until needed
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@edbiology Thanks for bringing up that point. Having the lid on keeps the agar solution protected from any dust or particles that could flow in. If you are working near a flame and are careful, you don't need to flame the actual cap and bottle. Flaming the cap/bottle is a good sterile technique and often done, but depending on your needs, it is not always necessary. Hope that helps =)
@nana22124 This video only shows how to pour the plates and not how to make the agar solution and that is why the autoclaving step is not shown.
When you make the agar solution, yes you need to autoclave it to sterilize the mixture. Then, if you are not using the mixture right away, the agar will solidify (so you can pre-make your agar mixtures, autoclave them, and store them in the solid state). Our video starts by using autoclaved solid agar, which you melt by microwaving.
Hope that helps!
Cool, thanks.
Thank you 🙏
ty
no flowhood?
I never use a flowhood or bio safety cabinet and have never had contamination problems, just pouring on the lab bench. If you are using antibiotics (amp, kan, etc) and keep the plates covered (except when pouring them of course) they will be fine. Just use basic clean techniques (gloves / lab jacket), don't breath or spit into the plates. I pour mine using serological pipettes which are sterile, instead of pouring directly from the flask.
+Steve Ayres how long does it take to use the serological pipettes to pour the plates? I guess it would create less bubles but even if you use 50 ml pipettes doesnt it take too much time and cause solidification of agar?
+Never Beaten It only takes a couple seconds per plate. I can get through 200ml of agar before it solidifies, no problem using 25ml serologicals with the speed on max.
I will give it a try then! thanks!! I never like the idea of pouring something heavy and contamination promoting, through a big bottle mouth.
is it necessary to autoclave it? in my bio class we had to autoclave it or just using a microwave is fine
It is indeed necessary to autoclave the agar medium before using it. Microwaving the media will fail to kill the endospores that might cause contamination later on as microwaving the media kills only the active vegetative cells.
No pressure cook/autoclave to kill bacteria and endospores? Wouldn't agar plates be contaminated if using this process? not sure what antibiotics you put in there. I would imagine it would have to be a very clean microwave not used for food
yea, this is a wet dream for trichoderma
Also there is controversy using antibiotic agar if its for a certain magic creature
Dima K
it depends... ive printed wilds with additional degree of success so i do use it in that capacity, otherwise.. i just keep making transfers. (shrug)
why is the antibiotic added??
Usually to test for antibiotic resistance in bacteria as a screen
Or because they're pouring unsterilized agar in open air without even flame sterilizing, so they need all the help the can get. lol
It is usually used when the plates are meant to be used for growing bacteria that are transformed with a plasmid with a resistance gene to the particular antibiotic. This would allow for selecting only the cells which have been transformed with the plasmid of interest.
Hello friends
How's biology going
@@chilla9697 k
Looks like bad technique--sorry--look at your bubbles
Your sterile procedure/technique fails on a massive scale!! This is meant to be a sterile process (vs an unsterile process you sterilize via autoclave/pc after making plates), a sterile process involves using sterilized Agar+pre-sterilized plates poured in a still air box or using a Laminar Flow Hood. Your burner is doing nothing ;but creating air movement of particulates/spores, never do this outside of a still air box or using a Laminar Flow Hood, the agar should also be sterilized in an autoclave/pc for 20-40 minutes (I go 40 myself to ensure sterilization) @15 psi then allowed to cool to about 55 degrees C (50ish is the hardening temp) to avoid condensation buildup (flipping the plates upside down for a few days allows moisture to be reabsorbed by the agar). Definitely going to need antibiotics following this TEK outside of the proper working environment. lol otherwise good video.