Description:
GenMute™
siRNA Transfection Reagent for Primary Keratinocytes is pre-optimized for transfecting
siRNA to primary keratinocytes. Primary
keratinocytes are known for notorious resistance for DNA and siRNA
transfections. With the pre-optimized transfection conditions, average 90%
silencing can be obtained.
Size & content:
-
GenMute™ Reagent,
1.0 ml, sufficient for ~833 reactions based on transfecting 20 pmoles
siRNA or miRNA mimics in 24-well plate
-
GenMute™ Transfection Buffer (5x ), formulated for maximal transfection efficiency, 8.0 ml
to make 40 ml working solution
Refer to the following optimal transfection conditions for maximal silencing
on primary keratinocytes. 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
~70%
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