can I autoclave agarose? - (Sep/18/2008 )
Hello, I have to add a mix of 0,5% low melting agarose plus IMDM media and FBS in a plate with CHO cells. I had some contamination problems so I wonder if I can autoclave the agarose... I think I will make the agarose ( 2%) in TAE buffer, then autoclave it. After autoclaving I will put the bottle in a 37°C bath. Is it a good idea? Or will I damage the agarose? Thank you!
Yes, it is just highly refined agar, so it can definitely be autoclaved.
Autoclaving is fine. 37 degrees holding is probably too low, as the agarose will be close to, if not under, its setting temperature.
If you want 0.5%, starting from 2%, you need to dilute 1 in 4, right? Even if you start with the agarose at 40-45C, it should remain liquid. Then, when you add the other components, the temperature will go down fairly quickly. I would add the most temperature-stable component first, and cause the temperature to drop by steps, so the most labile components are at an elevated temperature for the shortest time.
Thank you all very very much!