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
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
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
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.

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