Description:
GenMute™ siRNA Transfection Reagent for MEFs is pre-optimized for transfecting siRNA to Mouse Embryonic Fibroblasts (MEFs). MEFs are required to support the growth of undifferentiated mouse or human ES cells, iPS cells, thus they are also called feeder cells. MEFs are isolated from mouse embryos and are used at their early passages. Before MEFs are used as feeder cells, they must be treated by irradiation or mitomycin C to stop the cells from further dividing.
Refer to the following optimal transfection conditions for maximal silencing on MEFs. GenMute™ reagent, 1.0 ml, is sufficient for ~833 transfections in 24 well plates or ~416 transfections in 6 well plates.
Summary of Optimal Transfection Conditions: Cell culture medium Confluency of cell on the day of transfection Optimal siRNA concentration Diluent for siRNA and GenMute™ Reagent Incubation Time to Form GenMute™/siRNA Complex Maximal Silencing Efficiency Transfection Results: Reporter Gene Silencing Efficiency (% ) | DMEM with 4.5 g/L glucose, 10% FBS ~50% 40 nM 1xGenMute™ Transfection Buffer ~15 minutes at RT 48 hours Renilla Luciferase 90% |
Storage:
Store at 4 °C. If stored properly, the product is stable for 12 months or longer.
Data Sheet & Protocol .pdf