Contents Per Kit:
● 1×1.0 ml of GenJet™ DNA Transfection Reagent
● 1×1.0 ml (x50) of HapiCell™ Transfection Toxicity Removal Cocktail
Description
GenJet™ DNA In Vitro Tranfection Reagent for SaoS-2 Cells is pre-optimized for transfecting human epithelial-like osteosarcoma cell line, SaoS-2. SaoS-2 (sarcoma osteogenic) are a non-transformed cell line derived from the primary osteosarcoma of an 11-year old Caucasian girl (since identified as Alice) in 1973 by Fogh et al[1]. In 1987, Rodan determined that Saos-2 cells “possess several osteoblastic features and could be useful as a permanent line of human osteoblast-like cells and as a source of bone-related molecules”. With our pripariotary transfection booster conjugation technology, this reagent was formulated to transfect Saos-2 cells with up to 60% efficiency (GFP positive cells).
SaoS-2 cell 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. Refer to the following optimal transfection conditions for maximal transfection efficiency on SaoS-2 cells. To achieve maximal efficiency, the “Shaved Cell Transfection” protocol was optimized for transfecting SaoS-2 cells. Refer to the following optimal transfection conditions for maximal transfection efficiency on SaoS-2 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%
1.3×10^5 cells per square centimeter
4:1
Serum-free DMEM with 4.5 g/L glucose
15 minutes at RT
20 minutes at 37 degree
48 hours
EGFP
pEGFP-N3 (CMV promoter)
60% |
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 SaoS-2 Cells. Saos-2 cells were grown per ATCC recommended culture medium and transfected with GFP cDNA (pEGFP-N3) by GenJet™. The efficiency was checked 24 hours post transfection by Nikon Elipse 2000 confocal microscopy
Data Sheet
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