293T Cell Passaging Protocol

From ZhangLabWiki
Jump to navigation Jump to search
  1. Add 1% anti-anti (in fridge) and 10% Fetal Bovine Serum (FBS) to DMEM media. Warm media in 37 C water bath.
  2. Suck out media from flask.
  3. Add 4 mL Trypsin to flask. Incubate at 37 C incubator for 5 minutes
  4. Add 6 mL media to quench Trypsin
  5. From the flask, add 2.5 mL to each 15 cm dish and 1 mL to each flask.
  6. Add media up to 25 mL per 15 cm dish and 15 mL per flask. Gently shake back and forth and side to side (never in a circle).