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In side-by-side cotransfection experiments with traditional reagents developed for plasmid delivery, only Thermo Scientific DharmaFECT Duo Transfection Reagent resulted in efficient cell delivery of both siRNA (assayed by target gene knockdown) or microRNA reagents, along with plasmid (assayed by level of reporter expression). DharmaFECT Duo is an excellent choice for experiments requiring cotransfection, such as RNAi rescue or utilization of reporter systems.
For transfection of plasmid DNA or shRNA, please select Thermo Scientific TurboFect Transfection Reagent. For transfection of siRNA or microRNA only, please select Thermo Scientific DharmaFECT 1 Transfection Reagent.
DharmaFECT Duo transfection reagent achieves reliable cotransfection results for a variety of experimental goals. The graph demonstrates specific modulation of Renilla luciferase (Rluc) gene:
Rluc expression is similar for Plasmid Alone and Plasmid+Control
Down-regulation of the Rluc as a result of miRNA mimic (plasmid + miRNA mimic)
Excellent knockdown of the Rluc by siRNA (Plasmid + siRNA)
The psiCHECK-2 vector (100 ng/well; Promega) with cloned microRNA recognition sites for miR-28, miR-95, miR-30d, and miR-105, respectively, were complexed alone or with RNAi reagents (10 nM) using DharmaFECT Duo (0.2 μL/well) in MCF-7 cells at 10,000 cells/well (96-well plate). The RNAi reagents were Thermo Scientific Dharmacon miRIDIAN Mimics (Negative Control, miR-28, miR-95, miR-30d, and miR-105, respectively) or siRNA (Renilla luciferase pool). Firefly and Renilla luciferase expression was assessed at 48 hours using the Dual-Glo Luciferase Assay System (Promega) and normalized to identically treated psiCHECK-2 empty vector.
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