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| Cell lines | Cell type | Species | Reagents |
| 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 |
| Transfection problem | Possible cause | Suggested 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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