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Prepare media and add 1.5 % agar before autoclaving it (15g per liter).
After autoclavation, cool the media in a 55 degree waterbath. Do not
allow
the solution to cool below this temperature as the agar gets solid
at that temperature. At this step, one can leave the agar/media mixture
for a long time.
Before plating, add proper antibiotics and glucose if needed. Pour
ca 25 ml per small agar plate (9 cm diameter) and ca 250 ml per big agar
plate (20cm diameter) and let it cool. Keep at room temperatur over night
and store at 4 degrees.
Plates without antibiotica can be stored up to 6 months, with ampicillin,
typically 4 weeks.
Hints:
Many antibiotica are destroyed at high temperatures, it is important
to let the media cool before adding antibiotica.
Agar melts at more than 100 degrees but gets solid at 55.
When pouring many plates, one might want to grab a pile of them, sequentially lifting the lowest lid and simply pour the agar media until it covers the bottom, then proceed with the next plate and so on. Pouring too little media may result in cracks in the surface and should be avoided.
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