Griffitts:Bacterial Staining

Solutions

 * LB Broth
 * 0.85% NaCl
 * 20 mM propidium iodide (in kit in freezer)
 * 3.34 mM SYTO 9 (in kit in freezer)

Procedure

 * Grow the bacterial cells to an OD600 of 0.5
 * If the OD600 is > 0.5, dilute appropriately
 * Pellet at 13,200 rpm for 1 minute
 * Resuspend in 1 mL 0.85% NaCl
 * Incubate 10 minutes
 * Pellet at 13,200 rpm for 1 minute
 * Resuspend in 500 μL 0.85% NaCl
 * Add 0.75 μL of 20 mM propidium iodide
 * Add 0.75 μL of 3.34 mM SYTO 9
 * Incubate in the dark at room temperature for 15 minutes
 * Proceed to fluorescent microscopy
 * PI (red) stains dead cells
 * SYTO 9 (green) stains live cells