Description
GenJet™ DNA In Vitro
Transfection 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