Cell lines have different properties and are sensitive or insensitive to different manipulations. This is certainly true for transfections and different cell lines require the development of an efficient transfection protocol to optimize efficient delivery while minimizing toxicity. Here we provide our esiRNA transfection protocols for some cell lines used in our laboratory. The values provided here can only serve as guidelines and a detailed optimization should be carried out in the individual laboratories. We recommend using an eGFP expressing cell line in conjunction with an eGFP and a control esiRNA to optimize the transfection protocol.

Cell line Origin Confluency Amount esiRNA Plate format Transfection method/reagent
R1/E129Sv Mouse ES15-25%50 ng96 wellLipofectamine 2000
E14Tg2a129Sv Mouse ES15-25%50 ng96 wellLipofectamine 2000
E14Tg2a129Sv Mouse ES15-25%20 ng 384-well Lipofectamine 2000
E14Tg2a129Sv Mouse ES15-25%500 ng 6-well Lipofectamine 2000
E14Tg2a129Sv Mouse ES15-25%7500 ng 10-cm Lipofectamine 2000
SW48Human Colorectal carcinoma20-30%40 ng96- wellOligofectamine
DLD-1Human Colorectal carcinoma20-30%50 ng96- wellLipofectamine 2000
HCT116Human Colorectal carcinoma20-30%50 ng96- wellOligofectamine
HCT116Human Colorectal carcinoma20-30%1200 ng6-wellOligofectamine
RKOHuman Colorectal carcinoma20-30%50 ng96- wellOligofectamine
HeLaHuman cervical cancer20-30%15 ng384-wellOligofectamine
HeLaHuman cervical cancer20-30%30 ng96-wellOligofectamine
HeLaHuman cervical cancer20-30%1000 ng6-well Oligofectamine
U2OSHuman osteosarcoma25-35%60 ng384-well Effecten
NIH3T3Immortalized mouse embryonic fibroblasts10%-20%50 ng384 wellLipofectamine RNAiMAX
NIH3T3Immortalized mouse embryonic fibroblasts10%-20%200 ng96-wellLipofectamine RNAiMAX
NIH3T3Immortalized mouse embryonic fibroblasts10%-20%1600 ng12-wellLipofectamine RNAiMAX
HEK293Human embryonic kidney20-30%25 ng384-wellEffectene