Freezingcellsinliquidnitrogen
Take off MediaTrypsinate with 1ml x2 Dulbecco A trypsinAdd 7ml MediaPipette up and down to distribute cells throughout media (i.e. not clumped together)Add media to sterile falcon tube (15ml = 1 flask, 50ml = 5 flasks)Spin down 1-2K, 5 minsTake off mediaResuspend pellet in 9ml FCS and 1ml DMSODistribute in 1ml aliquots (10 cryovials)Move cells to -84oC overnight wrapped in cotton wool in a polystyrene boxFinally free......閱讀全文
Freezing cells in liquid nitrogen
Take off MediaTrypsinate with 1ml x2 Dulbecco A trypsinAdd 7ml MediaPipette up and down to distribute cells throughout media (i.e. not clumped togethe
Removing cells from liquid nitrogen
Put cryovial straight from storage and float in the 37篊 water bath- caution should be taken as on rare occasions vials can explode when heated up due
Freezing Cells
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
Freezing and Thawing cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Routine Splitting and freezing of cells
1. Grow cells to subconfluence in a flask. 2. Harvest as per normal and count. 3. Spin down 5min 1.2K in benchtop. Resuspend at 1.0 X 106/ml in 10%
Freezing and Thawing of Mammalian Cell Lines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.
Tissue Culture Methods-1
I. TYPES OF CELLS GROWN IN CULTURE Tissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often
Cell Thawing/Cell Freezing Protocol
Freezing Cells:Cells should be growing well or known to be in log phaseCount, collect and pellet cells in a 15mL test tubeResuspend in freezing media
Cell Thawing/Cell Freezing Protocol
Freezing Cells: Cells should be growing well or known to be in log phase Count, collect and pellet cells in a 15mL test tube Resuspend in fr
Cryopreservation of cell cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial con