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Figure 1. GAPD mRNA levels were quantified using real-time RT-PCR in the cells transfected with siRNAs targeting GAPD or non-silencing siRNA. Forty-eight hours post-transfection, the cells were harvested and analyzed by real-time RT-PCR for GAPDH mRNA expression levels. Data were normalized against the 18S rRNA signal. Control samples were either mock-transfected or untreated. Values are normalized to untreated sample. Data are means ± SD (n=3).
Gene silencing by RNA Interference (RNAi) is a powerful research tool for studying gene function in mammalian cells. RNAi is a biological phenomenon by which double stranded RNA (dsRNA) specifically reduces gene expression of its corresponding gene. Potent inhibition of specific gene expression is experimentally achieved by the transfection of small interfering RNA (siRNA), which binds RISC complex and cause degradation of target complementary mRNA molecules in the cell. Therefore, successful, potent RNAi experimentation is dependent upon the highly efficient delivery of the siRNA into cells by transfection of stable and functional siRNA molecules. Most DNA transfection reagents are incompatible with siRNA, sensitive to serum, or are capable of inducing cytotoxicity, which negatively affects gene expression studies. Transfection Reagents have been developed and optimized specifically for use with siRNA transfection and allow transfection in the presence of serum without lowering transfection efficiency or reducing cell viability. Ready-to-run transfection protocols eliminate the need for extensive siRNA optimization experiments. Altogen Biosystems kits are also compatible for transfection of microRNA (miRNA) molecules.
Article by Dr.Lakshmanan "A549 Transfection Reagent From Altogen Biosystems": Link
Altogen Biosystems is a life sciences company dedicated to the development, marketing and manufacture of cell type specific transfection reagents. Efficient delivery of DNA, RNA, siRNA, and microRNA enabled by advanced formulation of reagents and peculiar design of protocols. Altogen Biosystems offers a complete transfection system for a broad range of cell lines. All reagents are functionally tested to be highly reproducible, serum compatible, induce low toxicity, and can be used for co-transfection experiments, and high throughput applications.
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Altogen Custom Services provide specialized biotechnology and pharmaceutical services, including - generation of stable cell lines, RNA Interference (RNAi) services, assay development, screening and transfection services. Generation of stably-expressing cancer cell lines and primary cells can be very expensive and time-consuming. Altogen Services offers generation of stable cell lines by transforming the cell line of choice to stably express gene of interest or shRNA-encoding construct. Cloning and shRNA expression services are available.
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