Comparison of RNA Transfection Reagents
Recently, a postgraduate at the Shanghai Public Health Clinical Research Center conducted an RNA transfection comparison using two transfection reagents, Turbofect transfection reagent (Thermo) and EntransterTM-R4000 (Engreen). The comparison is as follows: experimental method Transfection reagents: Turbofect transfection reagent (Thermo) and EntransterTM-R4000 (Engreen Biosystem Co, Ltd) Cells to be treated: human CD8+ T cells 1. Method for transfection of Turbofect The culture volume per well was 100 μL, and the number of cells was about 105. Take 0.5 μL of agomir, add 9.5 μL of serum-free RPMI1640, and mix well; 0.2 μL of Turbofect transfection reagent and agomir dilution were mixed well. The preparation of the transfection complex was completed; after mixing, incubate for 15-20 minutes at room temperature, directly take 10 μL into the wells of the paved cells, continue to culture for 24 h, collect the cells, and wash with PBS for more than 3 times for RNA extraction. Preparation of transfected Mix: 100nMago/NC: (0.5μL agomir+0.2μLTurbofect+9.3μL serum-free RPMI1640)×2.5=1.25+0.5+23.25 2.EntransterTM-R4000 transfection The culture volume per well was 100 μL, and the number of cells was about 105. Take 0.5 μL of agomir, add 9.5 μL of serum-free RPMI1640, mix well, and make 10 μL of agomir dilution; Take 0.25 μL EntransterTM-R4000, then add 9.75 μL of serum-free dilution liquid, mix well to make 10μLEntransterTM-R4000 dilution; The EntransterTM-R4000 dilution and the agomir dilution were thoroughly mixed (can be blown 10 times or more with an oscillator or a sampler) and allowed to stand at room temperature for 15 minutes. Transfection of the complex is completed; Add 20 μL of the transfection complex to the cell suspension in the well, move the culture dish back and forth, and mix well; The cells were observed 6 h after transfection, and the medium was changed. After 24 h of culture, the cells were harvested and washed with PBS three times or more for RNA extraction. Preparation of transfected Mix: 100 nMago/NC: (0.5 μL agomir + 9.5 μL serum-free RPMI 1640) × 2.5 = 1.25 + 23.75; Etranster dilution: (0.25 μLEtranster + 9.75 μL serum-free RPMI 1640) × 5 = 1.25 + 48.75 ( After standing at room temperature for 5 minutes), 20 μL was taken into anago and an NC tube, and allowed to stand at room temperature for 15 minutes. Experimental result Conclusion: Entrans BioTM's EntransterTM-R4000 (ago/NC: 422912-fold) transfection reagent is superior to Turbofect transfection reagent after 24 h of transfection, based on the expression of the target miRNA expression level. (ago/NC: 285870 times). Discussion: From the experimental results, Engreen Biosystem's Entransten-R4000 (ago/NC: 422912 times) transfection reagent was superior to Turbofect transfection reagent (ago/NC: 285,870 times). EntransterTM-R4000 is a newly developed transfection reagent for RNAs such as siRNA, microRNA, mimic, inhibitor, mRNA and shRNA synthesized by Engreen Biosystem. EntransterTM-R4000 not only transfects small RNAs, but is also optimized for long-chain RNAs such as mRNA. The reagents can be used to introduce RNA into a variety of cell lines, including primary and suspension cells. High transfection efficiency can be achieved with or without serum and antibiotics. Uv absorbers are the most widely used light stabilizers, which can be divided into salicylate, benzophenone, benzotriazole, substituted acrylonitrile and triazines according to their structure. The most widely used light stabilizers are benzophenone and benzotriazole in industry. Quench agent is mainly metal complex, such as divalent nickel complex, and often used with ultraviolet absorber, play a synergistic effect. Imported Premium Uv Absorber,Latest Biochemical Synthesis,Premium Phenyl Salicylate,Value-Added Plastic Solutions Xingbang High Molecular Materials Co., Ltd. , https://www.chemicaladditive.com