Figure 3

Performance of the modified TRIzol protocol assessed by total protein extract protein recovery (mg mL^-1). To compare the performance of the modified TRIzol protocol versus the TRIzol^® manufacturer’s, mirVana^TM PARIS^TM kit and standardized laboratory (A+2X Buffers) protocols, 10⁶ HCT116 cells were plated in 14 independent 100 mm plates (one per experimental setting: lanes 1 to 14), and cells were processed for total protein extraction 48 h after plating. Protein extracts were processed in 100 μl of extraction buffer (samples 1–4), or resuspended in 100 μl of buffer (samples 5–14). Modified TRIzol protocol allowed up to ~50% recovery of total protein content in comparison with mirVana^TM PARIS^TM kit and standardized laboratory (A+2X Buffers), but importantly allowed a much higher protein recovery compared to the TRIzol manufacturer`s protocol, whose recovery yield was not suitable for downstream applications.