Description
GenJet™ DNA In Vitro Tranfection Reagent for COS Cells is pre-optimized for COS cell transfection. COS is a cell line often used by biologists when studying the monkey virus SV40. It is also often used to transfect cells to produce recombinant proteins for molecular biology, biochemistry, and cell biology experiments. Two forms of COS cell lines commonly used are COS-1 and COS-7. The COS cell line was obtained by immortalizing a cell line derived from kidney cells of the African green monkey with a version of the SV40 genome that can produce large T antigen but has a defect in genomic replication.
Refer to the following optimal transfection conditions for maximal transfection efficiency on both COS-1 and COS-7 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 50 to 100 transfections in 6 well plates.
Summary of Optimal Transfection Conditions:
Confluence on the day of transfection
Cell culture conditions
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA Complex
Presence of Serum/Antibiotics during Transfection
Change Medium 5 Hours After Transfection
Maximal Efficiency
Transfection Results:
Reporter Gene
Plasmid
Efficiency (GFP %)
|
~90%
DMEM with 4.5 g/L glucose, 10% FBS
3:1
Serum-free DMEM with 4.5 g/L glucose
15 minutes at RT
Yes
No
48 hours
EGFP
pEGFP-N3 (CMV promoter)
88%
|
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
Two Pictures Showing Transfection Efficiency of GenJet™ Reagent on COS-1 (upper panel) and COS-7 (lower panel) Cells Respectively. COS-1 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