Trizol RNA isolation - (Dec/02/2005 )
I'm trying to isolate RNA from dendritic cells using Trizol. Currently I' lysing the cells in Trizol, incubate 5 min at RT, add 200 uL chloroform, incubate, centrifuge and take the upper colourless phase to continue with. However, I'm suspecting a DNA contamination. I found somewhere that in that case you can better centrifuge directly after adding the Trizol to pellet the DNA and afterwards add the chloroform to the supernatant and continue with the protocol. Can anyone comment on this?
i think for recovery and proper dissociation of proteins/nucleic acids it's better to let mixture stands. So I would better follow the standard procedure but after precipitation of nucleic acids, DNase treating your sample.