We have the most complete collection of transfection reagents with exceptional performance for the delivery of DNA, siRNA, Invitrogen Stealth RNAi and Invitrogen BLOCK-iT RNAi Vectors, in traditional or difficult-to-transfect cell lines. Remember to refer to the Seven Steps to RNAi Success when planning RNAi Experiments.

General transfection protocols

IMPORTANT NOTE: We are currently having technical issues with our PDF downloads on this page, we are very sorry for the inconvenience. While we work to get them restored, please reach out to techservices@thermofisher.com with the cell line and product you are interested in and they will email you a PDF. 

Cell-type Specific Transfection Protocols for Stealth RNAi and siRNA

Cell Type Source Lipofectamine 2000 Lipofectamine RNAiMAX Oligofectamine
3T3L1 Mouse Fibroblast Protocol    
786-O Human renal cell adenocarcinoma cells   Protocol  
A549 Human lung carcinoma Protocol Protocol  
D3 Mouse Embryonic Stem Cells Protocol    
HCT116 Human colon carcinoma Protocol Protocol  
HEK293 Human embryonic kidney Protocol Protocol  
HeLa Human cervical carcinoma Protocol Protocol Protocol
HepG2 Human liver carcinoma   Protocol  
MDA-MB-435 Human ductal carcinoma cells   Protocol  
HMVEC-L Human lung microvascular endothelial cells Protocol  


HT1080 Human fibrosarcoma   Protocol  
Human Mesenchymal Stem Cells Human bone marrow   Protocol  
HUVEC Human umbilical vein endothelial cells   Protocol Protocol
MCF7 Human breast cancer   Protocol  
ME-180 Human cervical carcinoma   Protocol  
NIH 3T3 Mouse embryonic fibroblast   Protocol  
NRK Rat normal kidney Protocol    
P19 Mouse embryonal Carcinoma Protocol    
SK-N-SH Human neuroblastoma   Protocol  
Rat1 Rat fibroblast cells Protocol    

Customers also found Lipofectamine RNAiMAX transfection reagent to work well in the additional cell types.

Stealth RNAi and siRNA transfection concentrations
The transfection concentration of a Stealth RNAi or siRNA duplex is determined by dividing the number of moles of siRNA used by the final volume of the transfection (i.e. starting medium volume + transfection mixture volume).Using a 24-well plate we typically transfect 0.5-5 pmol of siRNA in a 100ul transfection mix to 500ul of medium in each well to give a 1 to 10nM final concentration.The certificate of analysis for Stealth RNAi or siRNA has instructions on making a 20µM stock solution. If transfecting in triplicate make 400µl of 5x transfection mix by adding 0.1–1 uL of the 20 µM stock to 399 µL of Gibco Opti-MEM. Then add 100µl of this transfection mix to each of your wells.20µM Stealth RNAi or siRNA = 20pmol/µL.

Cell line database
We provide many free cell line-specific transfection protocols in the Cell Line Database. The database is intended to enable researchers to easily find references related to the use of our products and cell lines of interest. This database is by no means exhaustive, but a collection of data dependent on the efforts of our Technical Service team.

Co-transfecting a vector and siRNA:
When co-transfecting plasmids and siRNA, use the transfection conditions appropriate for a plasmid transfection. The siRNA component usually adds negligible nucleic acid mass and can be ignored. Find more about the transfection conditions and references for your cell line in the Cell Line Database.

Troubleshooting transfection experiments
We've created a general table to help troubleshoot transfections using cationic lipids. In addition we have compiled a list of frequently asked questions about transfections.