Stable Cell Line Selection Suspension Cells

The transfection method determines the cell type for stable integration.

Stable cell line selection suspension cells.

It is known that liposome reagents can be used to transfer dna into adherent cell lines. My cells normally are shaken. The next day replace growth medium with selection medium supplemented with a range of antibiotic concentrations. How to make a stable cell line using suspension cells mar 21 2005 hi everyone.

A no antibiotic control is included. While viral methods or electrotransfection are used to deliver dna into primary cells or notoriously difficult to transfect suspension cell lines. I m trying to do the selection to get to a stable population. By selecting for stable integrants utilizing a selectable marker such as puromycin resistance a population of successfully transduced cells can be found.

2 5 5 0 x 10 5 cells ml. I m working with a suspension cho cell line and have transfected a plasmid containing zeocin resistance. Maintain each concentration in triplicates. The first critical step for stable cell line generation is determining the optimal antibiotic concentration for selecting stable cell colonies.

A kill curve is a dose response experiment where the cells are subjected to increasing amounts of antibiotic to determine the minimum antibiotic concentration that is needed to kill all the cells over. Well i would say that culturing cells in the media qsupplemented with selection antibiotic would give you good results for establishing cell line.