GenJet™ In Vitro DNA Transfection Reagent for MEFs

Price: $178.00

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  • Catalog #: SL100489-MEF
  • Unit Size: 1.0 ml

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Description

GenJet™ DNA In Vitro Transfection Reagent for MEFs is pre-optimized and conditioned for transfecting Mouse Embryonic Fibroblasts (MEFs). MEFs are required to support the growth of undifferentiated mouse or human ES cells, iPS cells, thus they are also called feeder cells.  MEFs are isolated from mouse embryos and are used at their early passages.  Before MEFs are used as feeder cells, they must be treated by irradiation or mitomycin C to stop the cells from further dividing.  However MEFs are nortoriously hard to transfect.  With the pre-optimized reagent, you will get up to 85% GFP+ MEFs.

T
he "Shaved Cell Transfection" protocol was optimized to achieve maximal transfection efficiency. Refer to the following optimal transfection conditions for maximal transfection efficiency on MEFs cells. GenJet™ reagent, 1.0 ml, is sufficient for 500 transfections in 24 well plates or 250 transfections in 6 well plates.

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 c
entimeter
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)

85%

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 MEFs. MEFs were grown per ATCC recommended culture medium and transfected with pEGFP-N3 by GenJet™. The efficiency was checked 24 hours post transfection

Data Sheet




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