SF cell line 4D-Nucleofector X kit S; a total of 20 μl of supplemented SF solution is used per transfection. Likewise, we recommend calculating for 20% more volume than required.24Remove the medium completely from the pelleted cells from Step 22, and gently resuspend the cells in an appropriate volume (20 μl per 2 × 105 cells) of S1-supplemented SF solution. Do not leave the cells in SF solution for extended periods of time.25Pipette 20 μl of resuspended cells into each DNA premix from Step 19. Pipette gently to mix and transfer to a Nucleocuvette strip chamber. Repeat this step for each transfection condition.26Electroporate the cells by using the Nucleofector 4D program recommended by Amaxa, CM-130.27Gently and slowly pipette 100 μl of warm D10 medium into each Nucleocuvette strip chamber, and transfer all the volume into a well with the prewarmed medium from Step 18.
