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    Transfection

    Reagents optimized for your specific application

    Dharmacon transfection reagents are designed specifically for small RNA transfection (DharmaFECT), co-transfection of plasmid and small RNA (DharmaFECT Duo), plasmid transfection (DharmaFECT kb) and self delivering siRNA for difficult to transfect cell types (Accell siRNA).

    One of the key success factors in any gene modulation experiment (RNAi, overexpression, gene editing) is the introduction of RNA and/or DNA components into your cell line or in vivo system. When applying lipid-based transfection reagents, there are three general measures of an effective transfection reagent:

    • High efficiency delivery
    • Low cellular toxicity
    • Broad dynamic range

    We offer a variety of Dharmacon DharmaFECT transfection reagents with demonstrated success for all of these key criteria. Not only do we specialize in transfection of small RNA (siRNA, microRNA, crRNA, tracrRNA) but we offer DharmaFECT Duo, specifically designed for co-delivery of small RNA with plasmid DNA, and DharmaFECT kb for delivery of plasmid DNA.

    • DharmaFECT 1 Transfection Reagent

      The most broadly applicable DharmaFECT formulation for optimal siRNA or microRNA transfection into a wide range of cell types for successful RNAi experiments. DharmaFECT 1 has been validated in over 35 cell types.

    • DharmaFECT 2 Transfection Reagent

      One of four siRNA/microRNA specific formulations, DharmaFECT 2 is a chemically distinct alternative to one-size-fits-all transfection reagents to achieve high-efficiency silencing in more cell types.

    • DharmaFECT 3 Transfection Reagent

      One of four siRNA/microRNA specific formulations, DharmaFECT 3 is a chemically distinct alternative to one-size-fits-all transfection reagents to achieve high-efficiency silencing in more cell types.

    • DharmaFECT 4 Transfection Reagent

      DharmaFECT 4 Transfection Reagent is chemically distinct from the other DharmaFECT formulations, providing an alternative when DharmaFECT 1 does not achieve optimal results, and is ideal to include in transfection optimization experiments prior to RNAi screening or other high-value experiments

    • DharmaFECT Set of 4 Transfection Reagents

      An aliquot of each of the four DharmaFECT formulations for siRNA/microRNA transfection optimization studies. Useful for determination of the best reagent for your cells and particular experimental conditions.

    • DharmaFECT Duo Transfection Reagent

      For high-confidence co-transfection

    • DharmaFECT kb Transfection Reagent

      Efficient transfection of plasmid DNA with minimal cytotoxicity

    • Accell siRNA Reagents

      Targeted gene silencing in difficult-to-transfect cells

    DharmaFECT Transfection Reagent

    The table below lists transfection recommendations to help you select the appropriate DharmaFECT formulation for your research. You can also download a PDF version of the DharmaFECT Cell Type Guide here.

    All experimental conditions resulted in cell viability and positive control gene silencing of 80% or better.
    All experiments were done in 96-well plates with Non-targeting control siRNA and PPIB (Cyclophilin B) or GAPD Control pools at 25 nM; Alamar Blue (viability) and knockdown measured at 24 hours.
    Data normalized to untransfected for viability and both untransfected and Non-targeting control for knockdown.
    Transfection conditions should always be re-evaluated in the context of a new plate format or assay-specific requirements for cell density.
    Human
    Cell line Cell type Recommended DharmaFECT formulation DharmaFECT volume/well (μL) in 96 well plate Plating density in 96 well plate (cells per well) Other successful formulations
    786-0 Kidney adenocarcinoma 1 0.4 5 × 103 2
    A549 Lung carcinoma 1 0.2 1 × 104 2, 3, 4
    BxPC3 Pancreas adenocarcinoma 2 0.2 5 × 103 1, 3, 4
    DLD-1 Colorectal adenocarcinoma 2 0.4 5 × 103 1,3
    DU 145 Prostate carcinoma 1 0.2 1 × 104 2, 3, 4
    NCI-H1299 Lung carcinoma 2 0.2 1 × 104 4
    HCT-116 Colorectal carcinoma 2 0.1 5 × 103 4
    HEK293 Kidney transformed embryonic cells 1 0.2 1 × 104 2, 4
    HeLa Cervical epithelial adenocarcinoma 1 0.2 5 × 103 2, 3, 4
    HeLa S3 Cervical epithelial adenocarcinoma 4 0.4 5 × 103 1, 2, 3
    Hep G2 Hepatocellular carcinoma 4 0.4 1 × 104 1, 2
    hMSC Mesenchymal stem cells 1 0.4 5 × 103 2, 3, 4
    HT-29 Colorectal carcinoma 1 0.2 5 × 103 2, 3, 4
    HT1080 Fibrosarcoma 4 0.2 5 × 103 1, 2, 3
    Huh-7 Hepatocarcinoma 4 0.05 5 × 103 1, 2
    HUVEC Umbilical vein endothelial cells 4 0.2 2 × 104 1, 2
    LNCaP Prostate carcinoma 3 0.2 1 × 104 1
    MCF-10a Breast adenocarcinoma 1 0.2 1 × 104 2
    MCF-7 Breast adenocarcinoma 1 0.2 1 × 104 2, 4
    MDA-MB-453 Breast adenocarcinoma 2 0.2 1 × 104 1, 3, 4
    MDA-MB-231 Breast adenocarcinoma 4 0.1 5 × 103 1
    OVCAR-3 Ovarian adenocarcinoma 1 0.1 5 × 103 2, 3, 4
    PC-3 Prostate carcinoma 2 0.2 1 × 104 3
    SK-BR3 Breast adenocarcinoma 2 0.2 1 × 104 1, 3, 4
    SK-OV-3 Ovarian adenocarcinoma 3 0.4 1 × 104 1, 2, 4
    u87MG Brain glioblastoma 1 0.1 5 × 103 2, 3, 4
    Rodent
    Cell line Cell type Recommended DharmaFECT formulation DharmaFECT volume/well (μL) in 96 well plate Plating density in 96 well plate (cells per well) Other successful formulations
    A7r5 Rat aortic smooth muscle 2 0.1 5 × 103 1
    C2C12 Mouse myoblasts 1 0.2 5 × 103 2, 3, 4
    CHO K1 Chinese hamster ovary cells 4 0.8 1 × 104 1, 2
    ES-D3 Mouse embryonic stem cells 1 0.2 2 × 103 2
    ES-E14TG2a Mouse embryonic stem cells 1 0.2 2 × 103 2
    H9c2 Rat myoblasts 1 0.2 1 × 104 2, 3, 4
    J774A.1 Mouse macrophages 4 0.2 1 × 104 -
    NIH/3T3 Mouse embryonic fibroblasts 1 0.2 1 × 104 3
    NRK-49F Rat kidney fibroblasts 2 0.2 1 × 104 1, 4
    Rat2 Rat fibroblasts 1 0.2 2 × 104 2
    3T3-L1 Mouse embryonic fibroblasts 1 0.2 5 × 103 3
    Other
    Cell line Cell type Recommended DharmaFECT formulation DharmaFECT volume/well (μL) in 96 well plate Plating density in 96 well plate (cells per well) Other successful formulations
    COS-7 African green monkey kidney 2 0.4 5 × 103 1, 3, 4

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