I am experiencing some problem ( actually I do not know if it is a problem) with RNA extraction technique.
I use the Trizol protocol. After separating the phases, I take the supernatant that is clear and (make sure that i donīt touch the tip in the interphase to avoid contamination) and I mix with isopropol alcohol. With centrifugation a pellet is formed in the bottom but, together with that, some very very small white pieces appear suspended in isopropol. Sometimes these pieces stay together resembling snow flakes.
At the beginnig I had decided not to discart these flakes and to keep together with the pellet. The amount of RNA and the 260/230 and 260/280 ratios was OK. But recently I have decided to discart these flakes and to keep just the pellet ( because i put in my mind that the flakes coundnīt be RNA), and the ratios continued OK but my amount of RNA decreased a lot! Could the flakes be RNA?
The sample is human gum. Iīve already swapped the Trizol, the chloroform and the isopropanol and nothing has changed.
Someone could help me?
0
Trizol RNA Extration problems purification step - snow flakes
Started by Lilip, Jan 25 2011 03:28 PM
No replies to this topic
