What are the suggested cell washing steps?
• After pipetting the entire sample above the gel layer into a separate tube, add sterile PBS (phosphate buffered saline) to bring volume to 15 mL. Cap the tube. Mix the cells by gently inverting the tube 5 times. • Centrifuge the tube for 15 minutes at 300 RCF. Aspirate as much supernatant as possible without disturbing the cell pellet. • Resuspend the cell pellet by gently vortexing or tapping tube with your index finger. • Add sterile PBS to bring the volume to 10 mL. Cap the tube. Mix the cells by gently inverting the tube 5 times. • Centrifuge the tube for 10 minutes at 300 RCF. Aspirate as much supernatant as possible without disturbing the cell pellet. Resuspend the cell pellet in the desired medium for the subsequent procedure.
