siRNA introduction to C2C12(Mouse muscle myoblast)

Cell Information

Cell condition: 2~10~5 cells/35mm dish(seeded)

Cells are differentiated in 1.5 mL of differentiation medium for 24 hr, and medium is replaced with Opti-Mem (non-serum) just before transfection

Medium: Multiplicated in:
DMEM + 10 fetal calf serum+2mM glutamine

Differentiated in: 
DMEM{0.1% fetal calf serum{5g/ml Transfferin{5g/ml insulin+2mM glutamine

Seeded and maintained in: 
1.5 mL of growth medium

Protocol

Reagent used: GenomONE-Neo
Trangient or Stable: ---
Introduced molecule: Bin-1 siRNA
Molecule conc.: 25M
Method: Method-2
Reagent volume: <Incorporation step>
HVJ-E: 20L
siRNA sol.: 15L
Reagent A:
Buffer:
Reagent B: 1.5L
<Introduction step>
Reagent C: 2.5L
Buffer: 32.5L
Treatment/well: 35L/well of 35mm dish with 1.5mL of Opti-Mem
Incubation time: After 4hr incubation, remove medium and add 1.5mL of fresh differentiation medium, and incubate for further 72hr.

Result

Efficiency: ---

Reference: Chie Kojima et al.@EMBO Journal, 23, 4413-4422 (2004).