Description
GenJet™ DNA In Vitro Tranfection Reagent for CV-1 is pre-optimized for transfecting CV-1 cells. The CV-1 cell line was initiated in March of 1964 with a tissue section excised from the kidney of an adult male African green monkey (Cercopithecus aethiops). The fibroblast line was originally utilized in research focusing on the transformation of the cancer-causing Rous sarcoma virus (RSV), but now is popular as a host for acquired immunodeficiency disease (AIDS) research, as well as transfection experiments with simian virus 40 (SV40) and recombinant plasmid vectors. CV-1 cells exhibit fibroblast morphology, grow adherently to glass or plastic surfaces, and are negative for reverse transcriptase. The cells are known to be susceptible to several viruses, including poliovirus 1, herpes simplex, simian virus 40, California encephalitis, and both Eastern and Western equine encephalitis. The cells exhibit rapid growth and chromosome number shifts have been reported to occur at high passage levels.
Refer to the following optimal transfection conditions for maximal transfection efficiency on CV-1 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 150 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 %)
|
~80%
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
36 hours
EGFP
pEGFP-N3 (CMV promoter)
75%
|
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 CV-1 Cells. CV-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