Lipid-based transfection reagents by Dharmacon are designed specifically for small RNA transfection (DharmaFECT 1-4), co-transfection of plasmid and small RNA (DharmaFECT Duo), and plasmid transfection (DharmaFECT kb).
One of the key success factors in any gene modulation experiment (RNAi, overexpression, gene editing or CRISPR modulation) 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
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.
Human |
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| Cell line | Cell type | Recommended DharmaFECT reagent |
DharmaFECT μL/well |
Plating density (cells/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 |
| ARPE-19 | Retinal pigment epithelial | 4 | 0.2 | 1 × 104 | 1,2 |
| 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 |
| Huh7 | Hepatocarcinoma | 4 | 0.05 | 5 × 103 | 1, 2 |
| HUVEC | Umbilical vein endothelial cells | 4 | 0.2 | 2 × 104 | 1, 2 |
| JEG-3 | Placenta, choriocarcinoma, epithelial | 3 | 0.2 | 1 × 104 | |
| LNCaP | Prostate carcinoma | 3 | 0.2 | 1 × 104 | 1 |
| MCF | Breast adenocarcinoma | 1 | 0.2 | 1 × 104 | 2, 4 |
| MCF 10A | Breast adenocarcinoma | 1 | 0.2 | 1 × 104 | 2 |
| MDA-MB-453 | Breast adenocarcinoma | 2 | 0.2 | 1 × 104 | 1, 3, 4 |
| MDA-MB-231 | Breast adenocarcinoma | 4 | 0.1 | 5 × 103 | 1 |
| OVCAR3 | Ovarian adenocarcinoma | 1 | 0.1 | 5 × 103 | 2, 3, 4 |
| PC-3 | Prostate carcinoma | 2 | 0.2 | 1 × 104 | 3 |
| Saos-2 | Bone, osteosarcoma, epithelial | 1 | 0.05 | 1 × 104 | 2,3,4 |
| SK-BR-3 | Breast adenocarcinoma | 2 | 0.2 | 1 × 104 | 1, 3, 4 |
| SKOV-3 | Ovarian adenocarcinoma | 3 | 0.4 | 1 × 104 | 1, 2, 4 |
| U-87 MG | Brain glioblastoma | 1 | 0.1 | 5 × 103 | 2, 3, 4 |
Rodent |
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| Cell line | Cell type | Recommended DharmaFECT |
DharmaFECT μL/well |
Plating density (cells/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 | 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 |
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| Cell line | Cell type | Recommended DharmaFECT |
DharmaFECT μL/well |
Plating density (cells/well) |
Other successful formulations |
| COS-7 | African green monkey kidney | 2 | 0.4 | 5 × 103 | 1, 3, 4 |
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.
Order DharmaFECT reagents
DharmaFECT 1
The most broadly applicable DharmaFECT formulation for optimal RNA transfection - validated in 35+ cell types.
DharmaFECT 2
A chemically distinct alternative to one-size-fits-all RNA transfection reagents - recommended for certain cell lines.
DharmaFECT 3
A chemically distinct alternative to one-size-fits-all RNA transfection reagents - recommended for certain cell lines.
DharmaFECT 4
A chemically distinct alternative to one-size-fits-all RNA transfection reagents - recommended for certain cell lines.
DharmaFECT set of 4
An aliquot of each of the four DharmaFECT formulations for determining the optimal reagent for your cells and experimental conditions.
DharmaFECT duo
For high-confidence co-transfection of small RNA and plasmid.
DharmaFECT kb
For efficient transfection of plasmid DNA with minimal cytotoxicity.
Helpful Resources
DharmaFECT Cell Type Guide
Choose Dharmacon™ DharmaFECT™ 1, 2, 3, or 4 for optimal transfection of siRNA or miRNA reagents.
Watch "Top tips for Transfection"
Use our tips for getting the best results out of your siRNA and CRISPR transfections.
Watch the DharmaFECT Duo video
Learn about the different reagents that can be co-transfected with ease using DharmaFECT Duo.
Inquire for special pricing on larger volumes
Contact usEditing of PSMD7 gene in U2OS-(Ubi)EGFP cells using Edit-R Cas9 Nuclease protein NLS delivered by DharmaFECT transfection reagents
U2OS-(Ubi)EGFP cells were plated at 10,000 cells/well in 96-well plates and co-transfected using DharmaFECT transfection reagents with 25 nM Edit-R Cas9 Nuclease protein NLS and synthetic crRNA:tracrRNA at 50 or 100 nM targeting PSMD7. Cells were harvested 72 hours post-transfection and the relative frequency of gene editing was calculated based on a DNA mismatch detection assay with T7 Endonuclease I. UT = untreated sample, MW = FastRuler Low Range DNA Ladder (Thermo Scientific).
Editing of VEGFA gene in HEK293T cells using Edit-R Cas9 Nuclease mRNA delivered by DharmaFECT transfection reagents
HeLa293T cells were plated at 20,000 cells/well in 96-well plates and co-transfected using DharmaFECT transfection reagents with 200 ng of Edit-R-Cas9 mRNA and synthetic crRNA:tracrRNA targeting VEGFA. Cells were harvested 72 hours post-transfection and the relative frequency of gene editing was calculated based on a DNA mismatch detection assay with T7 Endonuclease I. DF1 = DharmaFECT 1, Duo = DharmaFECT Duo, UT = untreated sample, MW = FastRuler Low Range DNA Ladder (Thermo Scientific).
DharmaFECT reagents provide an improved dynamic range
DharmaFECT reagents are effective across a broader range of experimental conditions when compared to Lipofectamine® 2000 (Invitrogen). Several cell densities and lipid volumes were investigated to determine optimal transfection conditions, shown by the shaded boxes. Three cell densities of HepG2 cells were transfected with GAPD siRNA (100nM) using a range of volumes (0.05 to 1.6μL/well) of Lipofectamine 2000 and DharmaFECT 4 transfection reagents. mRNA levels (bars) were assessed by branched DNA assay (Panomics Quantigene® Reagent System) and cell viability (data points) was determined by alamarBlue® (Biosource International).
DharmaFECT Transfection Reagents provide highly efficient delivery at low siRNA concentrations
To determine transfection efficiencies at low siRNA concentrations, two genes were targeted with various amounts of SMARTpool siRNA reagent. DharmaFECT achieved > 80% silencing at all siRNA concentrations, while HiPerFect (Qiagen) only achieved this level with PPIB at 100 nM. HeLa cells were transfected with SMARTpool TM reagents targeting Cyclophilin B (PPIB) or MAPKI at concentrations of 1, 5, 25, and 100 nM. mRNA expression (bars) was determined by branched DNA assay (Panomics Quantigene® Reagent System) and cell viability (data points) was determined by alamarBlue® (Biosource International).
DharmaFECT outperforms other reagents in optimization study
Table adapted from Borawski et al. A study by scientists at Novartis concluded that in transfection optimization in 384-well format (in preparation for screening) one of the four DharmaFECT transfection reagents outperformed all other reagents in delivery efficiency and overall cell viability in 9 of 10 cell lines. Lipid transfection reagents tested were DharmaFECT 1-4, HiPerFect® (Qiagen), TransIT-TKO® (Mirus) Lipofectamine® 2000 and Oligofectamine® (Invitrogen). J. Borawski et al., Optimization Procedure for small interfering RNA Transfection in a 384-well format. J. Bimolecular Screening,12, 546-559 (2007).
Choose from positive control siRNAs or our novel transfection indicators
| siGLO Positive Control Reagents | Species | Catalog Number |
|---|---|---|
| siGLO Cyclophilin B Control siRNA | Human, Mouse, Rat | D-001610-01 |
| siGLO Lamin A/C siRNA | Human, Mouse, Rat | D-001620-01 |
| siGLO Lamin A/C Control siRNA | Human | D-001620-02 |
| siGLO Lamin A/C Control siRNA | Mouse | D-001620-03 |
| siGLO Lamin A/C Control siRNA | Rat | D-001620-04 |
| siGLO Negative Control Reagents | Species | Catalog Number |
| siGLO RISC-Free Control | Human, Mouse, Rat | D-001600-01 |
| siGLO Transfection Indicators | Species | Catalog Number |
| siGLO Green Transfection Indicator | Human, Mouse, Rat | D-001630-01 |
| siGLO Red Transfection Indicator | Human, Mouse, Rat | D-001630-02 |
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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.
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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.
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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.
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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
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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.
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DharmaFECT Duo Transfection Reagent
For high-confidence co-transfection
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DharmaFECT kb Transfection Reagent
Efficient transfection of plasmid DNA with minimal cytotoxicity