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
|
~90% |
| Cell culture conditions |
DMEM with 4.5 g/L glucose, 10% FBS |
| GenJet™ (µl) : DNA (µg) Ratio |
3:1 |
Diluent for DNA and Transfection Reagent
|
Serum-free DMEM with 4.5 g/L glucose
|
| Incubation Time to Form GenJet™/DNA Complex |
15 minutes at RT |
| Presence of Serum/Antibiotics during Transfection |
Yes |
| Change Medium 5 Hours After Transfection |
No |
| Maximal Efficiency |
48 hours |
Transfection Results:
Reporter Gene
|
EGFP |
Plasmid
|
pEGFP-N3 (CMV promoter)
|
| Efficiency (GFP %) |
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
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