Maintenance tips
Maintenance of btl-3 cell line 1. Subculture no more than 1:4 2. Feed cells by replacing 90% of media every 3-4 days. 3. Only subculture when cells are 95% confluent. 4. To subculture, remove all media, save 10% of media. 5. Rinse with 0.5 mL Trypsin LE 6. Rinse again with 0.5 mL of Tryspin LE to cover the monolayer, and incubate for 4-5 minutes. 7. Add the appropriate volume of new media and the remaining old media to the cells. 8. Wash cells off the growing surface. 9. Pellet cells and resuspend using P1000 to promote single cell suspension. 10. Subculture to new flask or plates accordingly.