Splitting Cells: Difference between revisions
m →Protocol: added the aspiration processes and changed wash cells twice to "once" |
added SOP hyperlinks |
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==SOP== | |||
*[[SOP-_Biosafety Cabinet|SOP- Biosafety Cabinet]] | |||
*[[SOP-_Cryogenic Materials|SOP- Cryogenic Materials]] | |||
*[[SOP-_Vacuum Pumps|SOP- Vacuum Pumps]] | |||
==Materials== | ==Materials== | ||
*Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM | *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 -/- | *PBS -/- | ||
*0.05% Trypsin | *0.05% Trypsin | ||
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==Protocol== | ==Protocol== | ||
#Warm PBS and Media in water bath | #Warm PBS and Media in water bath | ||
# Aspirate media | # Aspirate the plate media | ||
#Wash cells once with 10 mL (per 10 cm dish) PBS -/- then aspirate the PBS | #Wash cells 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 1 mL trypsin and allow to sit in the hood for 2-5 min | ||