Contents Per Kit:
● 1×1.0 ml of GenJet? DNA Transfection Reagent
● 1×8.0 ml (5x ) of GenJet? DNA Transfection Buffer
Description
GenJet? DNA In Vitro Tranfection Reagent for HUVEC is pre-optimized for transfecting human umbilical vein endothelial cells (HUVEC). HUVEC is one of hard-to-transfect cell lines partly due to the transfection toxicity related cell death. In combination of our proprietary HapiCell? Toxicity Removal Cocktail, the pre-optimized transfection reagent gave up to 60% efficiency. To achieve maximal efficiency, the “Shaved Cell Transfection” protocol was optimized for transfecting HUVEC. Refer to the following optimal transfection conditions for maximal transfection efficiency on HUVEC. GenJet? reagent, 1.0 ml, is sufficient for 300 transfections in 24 well plates or 150 transfections in 6 well plates.
Summary of Optimal Transfection Conditions:
Cell culture medium
DMEM with 4.5 g/L glucose, 10% FBS
Confluency of cell on the day of shaving
~90%
Cell density for plating
1.3×10^5 cells per square centimeter
GenJet? (μl) : DNA (μg) Ratio
2.25:1
Diluent for DNA and Transfection Reagent
GenJet? DNA Transfection Buffer (1x )
Incubation Time to Form GenJet?/DNA Complex
15 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 %)
60%
Storage Condition Store at 4 °C. If stored properly, the product is stable for 12 months or longer Data Sheet
