Stable Cell Line Generation Protocol

Stable cell line generation is made possible by the use of positive selection markers such as hygromycin g418 geneticin zeocin and blasticidin antibiotic resistance.

Stable cell line generation protocol.

Unlike the short term protein expression observed using transient transfection methods generating cell lines using lentiviral vectors enables long term protein expression studies. Stable expression can be influenced by the transfection method used. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector. Protocol for generation of stable cell lines 2.

Major challenges for generation of stable cell lines are low transfection efficiency and or integration frequency. Adherent mammalian cells are sensitive to concentrations of 2 5 μg ml while cells in suspension are sensitive to concentrations as low as 0 5 2 μg ml. While common transfection methods such as lipofection can be used for the stable expression in easy to transfect cell lines e g hela cos 7. Puromycin has a fast mode action causing rapid cell death even at low antibiotic concentrations allowing the generation of puromycin resistant stable cell lines in less than one week.

The transfection method determines the cell type for stable integration. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies. As each stable cell line generation project is unique our service is tailored to meet your requirements and to ensure a safe investment. So far generation of stable cell lines has been a major challenge for many cell types e g jurkat mcf7 or u937 since overall transfection efficiencies and or integration frequencies have been low.

Selection markers can be delivered using the same plasmid that contains the gene of interest in cis or on a separate plasmid in trans that needs to be co transfected with. It is known that liposome reagents can be used to transfer dna into adherent cell lines. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector. Our xtencho tm cell line together with our in house xten protocol is your best chance at overcoming your.

Send your requirements to our phd account managers who will put together a customized offer. Protocol of stable cell line generation 1. Stable cell line generation.