docs/Protocols/Subculturing Cells.md

Before You Begin

• Sterilize everything with 70% ethanol.
• Spray your hands after touching anything outside the hood.

Protocol

1. Aspirate old media.
2. Wash with PBS, aspirate.
3. Add 1 ml Tryp-LE, incubate 5–10 min.
4. Dislodge cells, add 9 ml complete media.
5. Seed 1 ml into a new flask with 9 ml complete media.
6. Incubate at 37°C with 5% CO₂.