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!

---