Generation of a stable cell line: A positive control plasmid works, but I cannot obtain stable clones with my construct of interest. What could be the reason?


One reason could be that your gene product is toxic to the cells.

We recommend testing the construct for expression of the gene of interest by transient Nucleofection® of your cells. If the proportion of expressing cells drops between 4 and 48 hours post Nucleofection®, this can be an indication of your gene product being toxic.

A different reason could be that the DNA is degraded. (Linearized) plasmid should always be checked on an agarose gel for integrity before Nucleofection®.

Research Areas:
Gene Expression