Isolated Nuclei DNA Staining

Nuclei for Cell Staining Analysis

Hypotonic Staining Buffer:

Sodium citrate tribasic dihydrate 10.0 mg
Triton® X-100 30.0 ul
Propidium iodide 5.0 mg
Ribonuclease A 0.2 mg
H20 100 ml
  1. Aliquot 1x106 cells into each tube.
  2. Centrifuge 200 to 400 xg for 5 - 10 minutes and aspirate supernatant without disturbing the pellet.
  3. Add 500 ul of the Hypotonic Staining Buffer to the pellet and gently mix well.
  4. Keep samples at 4°C protected from light for 30 min or for a maximum of 1 h before acquisition on the flow cytometer.

Note:

Debris can increase with longer staining incubation times.

Reference

Krishan A: Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J. Cell Biol. 66:188-193, 1975.