Description
GenJet™ DNA In Vitro
Transfection Reagent for MDCK cell is pre-optimized for transfecting MDCK cells.
MDCK cell line is derived from the kidney of an apparently normal, adult, female
cocker spaniel; used in numerous viral studies; susceptible to vesicular
stomatitis, vaccinia, and Coxsackie viruses, and certain reoviruses and
adenoviruses.
The "Shaved Cell
Transfection" protocol was optimized to achieve maximal transfection efficiency. Refer to the following optimal transfection conditions for maximal transfection
efficiency on MDCK cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 transfections in 24 well plates or 150 transfections in 6 well plates.
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Summary of
Optimal Shaved CellTransfection Conditions:
Cell culture
medium
Confluency
of cell on the
day of shaving
Cell density for plating
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA
Complex
Incubation time of shaved cells with transfection complex
Maximal Efficiency
Transfection
Results:
Reporter Gene
Plasmid
Efficiency (GFP %) |
DMEM with 4.5 g/L glucose, 10% FBS
95~100%
1.3x10^5 cells per square centimeter
4:1
Serum-free DMEM with 4.5 g/L glucose
12 minutes at RT
20 minutes at 37 degree
48 hours
EGFP
pEGFP-N3 (CMV promoter)
70% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
A Picture Showing Transfection Efficiency of GenJet™ Reagent on MDCK Cells. MDCK cells
were grown per ATCC recommended culture medium and transfected with pEGFP-N3 by
GenJet™. The efficiency was checked 24 hours post transfection
Data Sheet 
