Dear Salman,

at first you need to have a plasmid encoding for your gene of interest and more importantly for a selection gene. Without any selection you will not be able to produce stably transfected cells.

Then you will need antibiotics for selection. there will be 3 steps for this:

Only after you can transfect your cells; you can wait 1 to 2 days before passaging them and culturing them in presence of antiobiotics.

The cells that have integrated the resistance gene will survive.

In order to perform efficient transfection, I would recommend you trying our DreamFect Gold transfection reagent. It presents a high compaction level and low toxicity and is ideal for the generation of stable transfected cells.

For your convinience you can find more information in the link below.

I will be glad to send you sample you DreamFect Gold, you can contact me via ResearchGate or directly at: [email protected]

Good luck with your experiments

Cedric

http://www.ozbiosciences.com/transfection-dna/19-dreamfect-gold-transfection-reagent.html

Please, take into consideration that a DNA vector, a transfection reagent, expression of an antibiotic resistance (trans)gene, expression of a reporter (trans)gene, and selection by acute/chronic antibiotic treatment may evoke cellular responses that affect the biochemical processes under investigation. In the following review, we demonstrate that an assumption that empty vector-transfected cells preserve the cytogenetic and phenotypic characteristics, and represent the adequate control in transfection experiments is not universally valid. We exemplify a number of studies, which reported obvious genomic, transcriptomic and phenotypic changes of tumor cells after transient/stable transfection of an empty vector. Finally, we conceptualize that the diverse experimental manipulations (e.g., transgene overexpression, gene knock out/down, chemical treatments, acute changes in culture conditions, etc.) may act as a system stress, promoting intensive genome-level alterations (chromosomal instability, CIN), epigenetic and phenotypic alterations, which are beyond the function of manipulated genes.

Please see the following manuscript for details:

Stepanenko AA, Heng HH. Transient and stable vector transfection: Pitfalls, off-target effects, artifacts. Mutat Res. 2017 Jul;773:91-103. doi: 10.1016/j.mrrev.2017.05.002.