Description
GenJet™ DNA In Vitro Tranfection Reagent for C2C12 Cell is pre-optimized for transfecting C2C12 cells. C2C12 cells were originally obtained by Yaffe and Saxel (1977) through selective serial passage of myoblasts cultured from the thigh muscle of C3H mice 70 h after a crush injury. These cells were shown to be capable of differentiation. C2C12 cells are a useful model to study the differentiation of non-muscle cells to skeletal muscle cells (e.g myosin phosphorylation mechanisms) and express muscle proteins and the androhen receptor (AR).
Refer to the following optimal transfection conditions for maximal transfection efficiency on C2C12 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 150 to 300 transfections in 6 well plates.
Summary of Optimal Transfection Conditions:
Cell culture conditions
|
DMEM with 4.5 g/L glucose, 10% FBS
|
Confluence on the day of transfection |
95~100% |
Cell density for plating
|
1.3×10^5 cells per square centimeter |
GenJet™ (µl) : DNA (µg) Ratio |
4:1 |
Diluent for DNA and Transfection Reagent
|
Serum-free DMEM with 4.5 g/L glucose
|
Incubation Time to Form GenJet™/DNA Complex |
12 minutes at RT |
Incubation time of shaved cells with transfection complex |
20 minutes at 37 degree |
Maximal Efficiency |
48 hours |
Transfection Results:
Reporter Gene
|
EGFP |
Plasmid
|
pEGFP-N3 (CMV promoter)
|
Efficiency (GFP %) |
65% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
Data Sheet