Stable Cell Line Selection Suspension Cells

It is known that liposome reagents can be used to transfer dna into adherent cell lines.

Stable cell line selection suspension cells.

My cells normally are shaken. The next day replace growth medium with selection medium supplemented with a range of antibiotic concentrations. 2 5 5 0 x 10 5 cells ml. How to make a stable cell line using suspension cells mar 21 2005 hi everyone.

Here is a protocol that may help you. A no antibiotic control is included. The first critical step for stable cell line generation is determining the optimal antibiotic concentration for selecting stable cell colonies. While viral methods or electrotransfection are used to deliver dna into primary cells or notoriously difficult to transfect suspension cell lines.

I m working with a suspension cho cell line and have transfected a plasmid containing zeocin resistance. Well i would say that culturing cells in the media qsupplemented with selection antibiotic would give you good results for establishing cell line. By selecting for stable integrants utilizing a selectable marker such as puromycin resistance a population of successfully transduced cells can be found. I m trying to do the selection to get to a stable population.

Maintain each concentration in triplicates. 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.