The phenomenon of RNA interference (RNAi) was first (unknowingly) observed when RNA was shown to inhibit protein expression in plants and fungi. In 1998, Fire and Mello first observed that double-stranded RNA (dsRNA) was the source of sequence-specific protein inhibition in C. elegans. This discovery prompted the RNAi mechanism as a tool to study gene function and for potential therapy. While the studies in C. elegans?were promising, RNAi was still limited in use to lower organisms because delivering long dsRNA polymers had many non-specific inhibitory effects in mammalian cells. However, further studies in plants and invertebrates demonstrated that dsRNA polymers are processed internally by an enzyme called Dicer into short double-stranded RNA oligomers, 21 to 22 nucleotides in length. These short Dicer cleavage products are the actual molecules that lead to RNAi, and were hence named short (or small) interfering RNAs (siRNA).
siRNA Oligos |
Custom siRNA Oligos |
GenePharma's siRNA manufacturing process is vigorously monitored and under strict quality control according to the ISO9000 Quality Standard System. Annealing dsRNA aliquots. |
Chemical Modified siRNA Oligos |
GenePharma's chemically modified siRNA oligos have a greater stability, longevity and effective time in cell culture and serum compared to standard siRNA. This makes them more suited to be used for in vivo?applications. |
siRNA Negative Control |
Negative controls are necessary in any siRNA experiment to rule out side effects caused by siRNA delivery. Customers can design their own siRNA negative control, or GenePharma can provide effective ready-made siRNA negative controls that have no homology with the target gene sequence. |
siRNA Positive Control |
Positive controls are also necessary to validate your experimental results and optimize your experimental procedures. By demonstrating efficient knockdown of a well-known constitutively expressed target gene you can show that transfection, RNA extraction and the detection method are reliable and have been successful. Available positive controls include Lamin A/C, GFP22, Luciferase GL2, MAPK1, Beta-Actin, Vimentin, P53, GAPDH and Cyclophilin B. |
siRNA Transfection |
RNAi Transfection Reagents |
GenePharma's siRNA oligos and transfection reagents are highly efficient and easy to operate. They are non-toxic to cells and can be stored at 4 ℃?for a long time. High transfection efficiency and good repeatability can be obtained even in culture media containing serum. Therefore, it is not necessary to change culture medium prior to transfection. |
RNAi Expression Vector |
shRNA Expression Vector |
The GenePharma shRNA Expression Vector has two enzyme cleavage sites: BamH I and BbsI. BbsI is a special restriction enzyme that produces an asymmetrical supplementary coherent terminal, which guarantees that the sequence is inserted in correct direction, and prevents the vector from self-circlizing. Our shRNA expression vectors are available with many kinds of selection markers to establish a stably transfected cell-line: Neo: Neomycin resistant gene; Hygro: Hygromycin B resistant gene; GFP: Neo: GFP reporter and the Kan/G418 resistant gene. A GFP reporter may help detect transfection efficiency and instruct RNAi action sites. |
RNA Extraction |
Ezol RNA Extraction Reagent |
Ezol performs well with small quantities of tissue (50-100 mg) and cells (5 × 106) as well as large quantities of tissue (≥1 g) and cells (>107) of human, animal, plant, or bacterial origin. The simplicity of the Ezol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. |
Enzol DNA Extraction Reagent |
Enzol performs well with small quantities of tissue (50-100 mg) and cells (5 × 106) as well as large quantities of tissue (≥1 g) and cells (>107) of human, animal, plant, or bacterial origin. The simplicity of the Enzol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. |
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RNAi Transfection and Control Kits |
RNAi-Startup GAPDH Control Kit: |
GenePharma's RNAi-Startup GAPDH Control Kit contains both GAPDH siRNA positive control and siRNA negative control. The kit can be used to optimize siRNA transfection conditions, but also as an internal control for your particular experiment. Fluorescent dye labeled dsRNA can be used directly to observe the transfection efficiency. By detecting GAPDH gene expression with Real-Time PCR one can monitor transfection efficiency. |
RNAi-Startup GAPDH Basic Control Kit |
GenePharma's RNAi-Startup GAPDH Control Kit contains both GAPDH siRNA positive control and siRNA negative control. The kit can be used to optimize siRNA transfection conditions, but also as an internal control for your particular experiment. Fluorescent dye labeled dsRNA can be used directly to observe the transfection efficiency. By detecting GAPDH gene expression with Real-Time PCR one can monitor transfection efficiency. The kit can be used together with Real-time PCR Core Reagent (Catalog Number QSG-070). |
RNAi Gene-Knockdown Easy Kit |
This kit contains all components for siRNA transfection, to evaluate the transfection efficiency and to verify RNAi knockdown efficiency with RT-PCR. The kit contains both GAPDH siRNA positive control and siRNA negative control, fluorescent dye labeled dsRNA and PCR amplification primers and probes. The transfection efficiency can hence be detected directly with fluorescent dye labeled dsRNA and with RT-qPCR. |
RNAi-Startup IFN Response Basic Control Kit |
In mammalian cells, long double stranded RNA can trigger an interferon (IFN) response thereby causing non-specific transcriptional inhibition. If you are unsure whether your siRNA sequence induces a non-specific IFN response, we recommend to use this kit to monitor non-specific IFN by Real-Time PCR. The kit can be used together with Real-time PCR Core Reagent (Catalog Number QSG-070). |
RNAi-Startup IFN Response Control Kit |
This kit offers extra control features if you are worried about non-specific IFN effects. In addition to components of the Basic Control Kit, this kit allows RT-PCR monitoring of PKR, OAS-1 and hStat1, three well known IFN-stimulated genes next to the GAPDH house-keeping gene. |
RNAi House-Keeping Gene Control Kit |
GenePharma has developed some convenient house-keeping gene control kits for all your RNAi experiments. Please see GenePharma Product Catalog Page C8 for more details. |
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