GenJet™ In Vitro DNA Tranfection Reagent for MEFs

¥ 1,780.00

购买数量:

  • 目录号: SL100489-MEF
  • 包装: 1.0 ml

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Description

GenJet™ DNA In Vitro Tranfection 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 Transfection Conditions:

Cell culture medium DMEM with 4.5 g/L glucose, 10% FBS
Confluency of cell on the day of shaving 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 %) 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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