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.
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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
