164
edits
Changes
added SOP hyperlinks
==SOP==
*[[SOP-_Biosafety Cabinet|SOP- Biosafety Cabinet]]
*[[SOP-_Cryogenic Materials|SOP- Cryogenic Materials]]
*[[SOP-_Vacuum Pumps|SOP- Vacuum Pumps]]
==Materials==
*Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM
(To prepare DMEM:
1 bottle of DMEM found in 4c in cell culture room
1 tube FBS found in -80 in cell culture room
5 mLs PSG found in door of 4c in cell culture room
500 mL bottle taken from lab
filter found on metal rack behind fume hood
•pour all ingredients into DMEM bottle
•screw filter onto 500 mL bottle
•attach hose onto arm of filter
•pour mixture into filter)
*PBS -/-
*0.05% Trypsin
==Protocol==
#Warm PBS and Media in water bath
# Aspirate the plate media#Wash cells twice once with 10 mL (per 10 cm dish) PBS -/-then aspirate the PBS#Add 1 mL trypsin and allow to sit in the hood for 2-5 min
#Add 10 mL media to each new dish
#Check cells for trypsinization, and if necessary tap the cells