We have the most complete collection of transfection reagents with exceptional performance for the delivery of DNA, siRNA, Stealth™ RNAi and 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

Cell-type Specific Transfection Protocols for Stealth™ RNAi and siRNA

Cell Type Source Lipofectamine® 2000 Lipofectamine® RNAiMAX Oligofectamine™
3T3L1Mouse FibroblastProtocol  
786-OHuman renal cell adenocarcinoma cells Protocol 
A549Human lung carcinomaProtocolProtocol  
D3Mouse Embryonic Stem CellsProtocol  
HCT116Human colon carcinomaProtocolProtocol 
HEK293Human embryonic kidneyProtocolProtocol 
HeLaHuman cervical carcinomaProtocolProtocolProtocol
HepG2Human liver carcinoma Protocol 
MDA-MB-435Human ductal carcinoma cells Protocol 
HMVEC-LHuman lung microvascular endothelial cellsProtocol 

 

HT1080Human fibrosarcoma Protocol 
Human Mesenchymal Stem CellsHuman bone marrow Protocol 
HUVECHuman umbilical vein endothelial cells ProtocolProtocol
MCF7Human breast cancer Protocol 
ME-180Human cervical carcinoma Protocol 
NIH 3T3Mouse embryonic fibroblast Protocol 
NRKRat normal kidneyProtocol  
P19Mouse embryonal CarcinomaProtocol  
SK-N-SHHuman neuroblastoma Protocol 
Rat1Rat fibroblast cellsProtocol  


“Customers also found Lipofectamine® RNAiMAX transfection reagent to work well in the additional cell types. Click here for details.

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 20µM stock to 399µl of 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 Invitrogen™ products and cell lines of interest. This database is by no means exhaustive, but a collection of data dependent on the efforts of Invitrogen™ Technical Service.

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.