Flow Cytometry to measure DNA content in Tetrahymena

Materials

 * Sucrose (1 M stock)
 * MgCl2 ( 1 M stock )
 * Triton X-100
 * Propidium Iodide (1 mg/mL stock)
 * 1×PBS

Method
Sign up for the flow cytometer before use.

Turn on the flow cytometer at least 20 minutes before using.
 * 1) Wash cells with PBS twice to remove culture medium.
 * 2) Lyse 100 mL of Tetrahymena cells (OD540 0.3 = 10^5 cells/mL). ) in PBS buffer containing 0.25 M sucrose, 10 mM MgCl2, and 0.5% Triton X-100.
 * 3) Add propidium iodide to the lysed cells at a final concentration of 20 µg/mL and allow the nuclei to stain for 1 hour prior to analysis using the C6 Flow Cytometer.