This distributes cells evenly about the plate.
Plat e cells transfection protocol.
After quickly thawing the cells in a 37ºc water bath immediately transfer the thawed cell suspension into a 15 ml tube containing 10 ml of culture medium.
Prepare complexes in the plate and directly add cells at twice the cell density as in the basic protocol in a 100 µl volume.
Establishing plat a cultures from frozen cells 1.
Surface areas may vary depending on the manufacturer.
High retroviral yields with plat e cells.
Cells will adhere as usual in the presence of complexes.
These cells require transfection of only an expression vector to produce retrovirus.
Transfect a platinum retrovirus packaging cell line such as plat a amphotropic or plat e ecotropic which stably expresses both mmlv gag pol and an amphotropic or ecotropic envelope protein with a retroviral expression vector.
Unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid based transfection protocols up to 50 or higher of cells can be transiently transfected.
Establishing plat e cultures from frozen cells 1.
You may perform rapid 96 well plate transfections by plating cells directly into the transfection mix.
Cloned genes can be transfected into cells for biochemical characterization mutational analyses investigation of the effects of gene expression on cell growth investigation of gene regulatory elements and to produce a specific protein.
After quickly thawing the cells in a 37ºc water bath immediately transfer the thawed cell suspension into a 15 ml tube containing 10 ml of culture medium.
18 24 hours prior to transfection plate phoenix cells at 1 5 2 million cells per 6 cm plate in producer cell growth media.
The platinum e plat e cell line a potent retrovirus packaging cell line based on the 293t cell line was generated using novel packaging constructs with an ef1α promoter to ensure longer stability and high yield retroviral structure protein expression gag pol ecotropic env.
They exhibit longer stability and produce higher yields of retroviralstructure proteins.
Transfection the delivery of dna or rna into eukaryotic cells is a powerful tool used to study and control gene expression.
After adding cells set plates in incubator and gently shake forward and backward then side to side 3 4 times.
