I am trying to figure out a protocol to separate cell debris from Jurkat cells using Percoll density centrifugation. Most protocols mention layering cells in PBS on top of stock isotonic Percoll (1:9, 10XPBS: Percoll, 1.17 g/mL). Can we instead mix the cells with stock isotonic Percoll & do the separation. Is there an pros/cons to layering cells in buffer on top vs. mixing them with Percoll.