Stable Transfection

* This product is for research use only. Not intended for use in the treatment or diagnosis of disease.

What is stable transfection?

Unlike transient transfection, stable transfection can integrate the transfected DNA into the cell genome and pass it to the progeny of the cell, thereby achieving the long-term introduction of foreign DNA into the cell. Stable transfection can be used to study the long-term effects of gene expression or to create cell lines with new characteristics for further experiments, such as screening potential drug candidates in discovery experiments.

How does stable transfection work?

The purpose of stable, long-term transfection is to isolate and propagate a single clone containing transfected DNA that has been integrated into the genome of the cell. One of the most reliable methods for selecting cells that stably express transfected DNA is selective screening. Commonly used selection markers include genes that are resistant to various selection drugs. Long-term continuous antibiotic treatment of cells will only lead to the expansion of stably transfected cells, and unstable cells will die due to a lack of antibiotic resistance.

An alternative strategy is to use vectors carrying essential genes that are defective in each cell line. For example, CHO cells that lack dihydrofolate reductase (DHFR) gene expression cannot survive without the addition of nucleosides. However, these cells, after being stably transfected with DNA expressing the DHFR gene, will synthesize the required nucleosides and survive. Another advantage of using DHFR as a marker is that when cells are exposed to increasing doses of methotrexate, gene amplification of DHFR and related transfected DNA occurs, resulting in multiple copies of the plasmid in the transfected cells.

Your stable transfection partner

We provide an unparalleled variety of transfection reagents to meet your different needs. For researchers who wish to establish stable protein expression in mammalian and insect cells in their laboratories, we provide variety types of stable transfection reagents to meet your different needs.

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DNA Transfection Kits
RNA Transfection Kits
Cell Line Transfection Kits

Tips for a stable transfection

  • Use high-quality reagents and purified plasmid DNA.
  • Optimize pDNA concentration and the ratio of transfection reagent to DNA.
  • Use fluorescently labeled DNA vectors (luciferase, GFP, b-gal).
  • Reduce serum concentration in cell growth medium during transfection.
  • Do not add antibiotics prematurely, wait at least 72-96 hours for the resistance gene to be fully expressed.
  • Experiments show that although the uptake of linear DNA is still lower than that using supercoiled DNA, linear DNA seems to be more suitable for stable transfection.