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Cells transfected successfully

Cell linesCell typeSpeciesReagents
3T3-F442A Adipocytes Mouse Lipofectamine Reagent 
SW13 Adrenal carcinoma Human Lipofectin Reagent 
Primary Amniocytes Human Lipofectin Reagent 
T24 Bladder carcinoma Human Lipofectin Reagent 
U-2 OS Bone sarcoma Human Lipofectamine 2000 Reagent 
KG-1; KG1a; K562 leukemia Human Lipofectamine 2000 Reagent 
SK-BR-3 Breast Cancer Human Lipofectamine 2000 Reagent 
MCF7 Breast Cancer Human Lipofectamine 2000 Reagent 
Hs 578T; T-47D; MDA-MB-231; ZR-75-1 Breast Cancer Human Lipofectamine Reagent 
C127 Breast Cancer Mouse Lipofectin Reagent 
NMU Breast Cancer Rat Lipofectamine Reagent 
HeLa Cervical Cancer Human Lipofectamine 2000, Lipofectamine Reagent 
C-33 A Cervical Cancer Human Lipofectamine Reagent 
HT-29 Colon Human cancer Lipofectamine 2000 Reagent 
HCT116 Colon Human cancer Lipofectamine 2000 Reagent 
COLO 320 HSR; SW40; 3SW480; Caco-2 Colon Human cancer Lipofectamine Reagent 
T84 Colon Human cancer Lipofectin Reagent 
L Connective Tissue Mouse Lipofectamine Reagent; Lipofectin Reagent 
Isikawa Endometrial cancer Human Lipofectamine Reagent 
Primary HUVEC Endothelial Human Lipofectin Reagent 
Primary HUAEC Endothelial Human Lipofectin Reagent 
Primary Endothelial Endothelial Rabbit Lipofectin Reagent 
P19; F9 Embryonic carcinoma Mouse Lipofectamine Plus Reagent 
Primary Fibroblast Fibroblasts Human Lipofectamine 2000 Reagent 
Mv 1 Lu Lung Fibroblasts Mink Lipofectin Reagent 
NIH 3T3; TA1; Y-2; NIH 3T6; 10T1/2 Fibroblasts Mouse Lipofectamine with Plus Reagent, Lipofectin Reagent 
MDCK Kidney Dog Lipofectamine Reagent 
BHK-21 Kidney Hamster Lipofectamine with Plus Reagent 
293, 293H, 293F Kidney Human Lipofectamine 2000 Reagent 
A498; UMRC 6 Kidney carcinoma Human Lipofectamine Reagent 
COS-1; COS-7; Vero CV-1 Kidney Monkey Lipofectamine 2000 Reagent 
LLC-PK Kidney Pig Lipofectamine Reagent 
FLK Kidney Sheep Lipofectamine Reagent 
Primary Liver Liver Dog Lipofectin Reagent 
Huh7 Liver cancer Human Lipofectamine 2000 Reagent 
Hep 3B Liver cancer Human Lipofectamine 2000 Reagent 
Primary Liver Liver Mouse Lipofectin Reagent 
Primary Liver Liver Rat Lipofectamine 2000 Reagent 
FTO-2B Liver cancer Rat Lipofectamine with Plus Reagent 
MSN610.2; Morris 7777; H35; McA-RH7777 Liver cancer Rat Lipofectamine Reagent; Lipofectin Reagent 
R1610 Lung Hamster Lipofectamine with Plus Reagent 
Dede Lung Hamster Lipofectamine Reagent 
MRC-5 Lung Human Lipofectamine 2000 Reagent 
WI-38 Lung Human Lipofectamine Reagent 
NCI-H23 Lung Human cancer Lipofectamine Reagent 
A549 Lung Human carcinoma Lipofectamine 2000 Reagent 
H187; NCI-H146 Lung Human small cell carcinoma Lipofectin Reagent 
LLC Lung Mouse carcinoma Lipofectamine Reagent 
BL3.1 Lymphoid Bovine leukemia Lipofectamine Reagent 
Jurkat Lymphoid Human lymphoma Lipofectamine 2000 Reagent 
U937 Lymphoid Human lymphoma Lipofectin Reagent 
MOLT-4 Lymphoid Human leukemia DMRIE-C Reagent 
A20.J; M12 Lymphoid Mouse B cell lymphoma Lipofectamine Reagent 
P388D1 Lymphoid Mouse lymphoma Lipofectamine Reagent 
THP-1 Macrophages/ Human monocyte Lipofectamine Reagent 
Primary; J774.16; RAW 264.7 monocytes Mouse macrophage Lipofectamine Reagent; Lipofectin Reagent 
B16 Melanoma Mouse Lipofectamine 2000 Reagent 
Trachea smooth muscle Muscle Cow Lipofectamine Reagent 
C2C12 Muscle Cow Lipofectamine with Plus Reagent 
Primary; G8 Muscle Mouse Lipofectamine Reagent 
Primary Muscle Rat Lipofectin Reagent 
L6E9 Muscle Rat Lipofectin Reagent 
A10 Smooth muscle Rat Lipofectamine Reagent 
U-87; A-172 Glioma Human Lipofectamine 2000 Reagent 
SK-N-MC Neuroblastoma Human Lipofectin Reagent 
SK-N-SH Neuroblastoma Human Lipofectamine Reagent 
Neuro-2a Neuroblastoma Mouse Lipofectamine Reagent 
Primary fetal astrocyte Astrocyte Rat Lipofectamine Reagent 
C6 Glioma Rat Lipofectamine Reagent 
PC 12 Pheochromocytoma Rat Lipofectamine 2000 Reagent 
CHO-K1 Ovary Hamster Lipofectamine 2000 Reagent 
CT60 Ovary Hamster Lipofectamine with Plus Reagent 
AS52; CHO-DXB11 Ovary Hamster Lipofectamine with Plus Reagent 
SKOV3 Ovary cancer Human Lipofectamine Reagent 
Primary Ovary Ovary Mouse Lipofectin Reagent 
Capan-2 Pancreas cancer Human Lipofectamine Reagent 
AR4-2J Pancreas Rat Lipofectamine Reagent 
Primary fetal islet cells Pancreas islet Rat Lipofectin Reagent 
AtT-20 Pituitary Mouse Lipofectamine Reagent 
Primary Pituitary Pituitary Rat Lipofectin Reagent 
GH3 Pituitary tumor Rat Lipofectamine Reagent 
LNCaP; PC-3 Prostate Human Lipofectin Reagent 
Primary keratinocytes Skin Human Lipofectamine 2000 Reagent 
AKR Spleen Mouse Lipofectin Reagent 
Sperm Sperm Mouse Lipofectin Reagent 
Primary hematopoietic CD34+ Stem cells Human DMRIE-C Reagent 
D3 Stem cells Mouse Lipofectin Reagent 
SK-UT-1 Uterus Human Lipofectamine Reagent 
Blastoderm cells  Chicken Lipofectin Reagent 
Primary chondrocytes  Chicken Lipofectamine Reagent 
Primary fibroblasts  Chicken Lipofectamine Reagent, Lipofectin Reagent 
Primary hepatocytes  Chicken Lipofectin Reagent 
BMS-M protoplasts  Corn Lipofectin Reagent 
Dictyostelium   Lipofectin Reagent 
A6 kidney cells Xenopus laevis Frog Lipofectamine with Plus Reagent 
Eggs; kidney cells Xenopus laevis Frog Lipofectin Reagent 
D.Mel-2; Schneider’s L2 Drosophila Insect Cellfectin Reagent 
Sf9; Sf21; High Five™ Spodoptera frugiperda Insect Cellfectin Reagent 
Neurospora crassa Neurospora crassa Insect Lipofectin Reagent 
DGZ protoplasts  Rice Lipofectin Reagent 
Primary lymphoid  Shrimp Lipofectin Reagent 
Protoplasts  Tobacco Lipofectin Reagent 
Saccharomyces cerevisiae Saccharomyces cerevisiae Yeast Lipofectin Reagent 
Schizosaccharomyces pombe Schizosaccharomyces pombe Yeast Lipofectin Reagent 

Troubleshooting

Transfection problemPossible causeSuggested solution
Low transfection efficiency. Not using optimal cationic lipid reagent. Select the cationic lipid reagent likely to result in the highest transfection efficiency for your cell type. See our selection guide
Low transfection efficiency. Not using optimal cationic lipid reagent concentration, DNA concentration, time or cell density. Optimize these parameters under final transfection conditions. See the cell-specific transfection protocols.
Low transfection efficiency. DNA-cationic lipid reagent complexes not formed. Do not use serum during complex formation step. Opti-MEM I Medium or D-MEM are good media for complex formation. If using serum containing medium for transfections, form the complex in the absence of serum. Note: Do not use Opti-MEM I Medium with Lipofectamine with Plus Reagent or for insect cells. Sf-900 II SFM gives optimal results for Sf9 or Sf21 cells. For D. mel-S2 cells, use Drosophila-SFM. 
Low transfection efficiency. Inhibitors were present. Do not use antibiotics, EDTA, citrate, phosphate, RPMI medium, chondroitin sulfate, hyaluronic acid, dextran sulfate, or other sulfated proteoglycans in the medium used to prepare complexes. 
Low transfection efficiency. Cationic lipid was frozen. Do not use cationic lipid reagents that have been frozen. Store at +4°C. 
Low transfection efficiency. Improper cell density. Cell density should be 70% to 90% confluent at time of transfection. (30%-70% confluent if using Optifect™ Transfection Reagent) 
Low transfection efficiency. Problems with the transfection assay. Include a positive control for the transfection assay. 
Low transfection efficiency. The promoter-enhancer of the transfected DNA is not recognized by the cell type. Make sure the transfected DNA is compatible with the target cell type. 
Low transfection efficiency. For Lipofectin Reagent, no preincubation of cationic lipid with medium. Preincubate Lipofectin Reagent in medium 30 min. prior to addition of diluted DNA. 
Low transfection efficiency. Diluted Lipofectamine 2000 Reagent was incubated too long before mixing with diluted DNA. For dilutions prepared in Opti-MEM I Medium, be sure that the diluted Lipofectamine 2000 Reagent is combined with the diluted DNA within 30 min. If D-MEM is used as the diluent, mix with the diluted DNA within 5 min. 
Low transfection efficiency. Cell density too low for Lipofectamine 2000 Reagent transfection. Use cells at 90% confluency for Lipofectamine 2000 Reagent transfections. 
High cell death (toxicity) Too much DNA. Perform a dose-response curve to determine the optimal amount of DNA. Include a cationic lipid reagent in the dose-response transfections, as DNA alone has a minimal effect on cell growth. 
High cell death (toxicity) Too much cationic lipid reagent. Perform a dose-response curve to determine the optimal amount of cationic lipid reagent. Include DNA with the cationic lipid reagent in the dose-response experiment, as cationic lipid reagent alone has a minimal effect on cell growth. 
High cell death (toxicity) Too few cells. Perform a dose-response curve to determine the optimal number of cells per transfection. Balance cell number with efficiency for your application. 
High cell death (toxicity) Cell viability decreased without serum. Use Opti-MEM I medium. Reduce or omit the number of washes in serum-free medium. Use 5 to 10% serum in the transfection medium. Be sure to form complexes in the absence of serum. 
High cell death (toxicity) For stable transfection, selection antibiotic added too soon. allow at least 48 h for cells to express resistance before adding selective antibiotic. 
High cell death (toxicity) Antibacterial agents were used during transfection. Do not use chloroquine, penicillin, or streptomycin during transfection because cationic lipid reagents make cells more permeable. 
Transfection not reproducible Confluency at the time of transfection varied. Keep transfection parameters such as confluency and phase of growth consistent. 
Transfection not reproducible Cells changed in culture. If possible, thaw fresh cells or obtain new cell line. 
Cationic lipid reagent solution is cloudy.  For Cellfectin DMRIE-C, and Lipofectamine 2000 Reagents this is normal. Be sure to mix reagent (invert 5 to 10 times) prior to taking an aliquot for transfection. 
Cationic lipid reagent solution is cloudy.  For Lipofectin the reagent may have been frozen or stored below +4°C. Do not use. 
Cationic lipid reagent solution is cloudy.  For Lipofectamine Reagent, after dilution in Opti-MEM I this is sometimes seen. This can be used for transfection. 
Precipitation of DNA-cationic lipid reagent complex. Excess EDTA present. Dissolve the DNA in water or in TE with the EDTA concentration <0.3 mm in the diluted DNA. 
Precipitation of DNA-cationic lipid reagent complex. Excess precipitation of cationic lipid reagent-DNA observed in complex. Ensure concentrations of cationic lipid reagent and DNA do not exceed recommended amounts in complex formation. 
Precipitation of lipid reagent. Reagent may have been frozen or stored below +4°C. Do not use. 
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