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A Modified Method for Transfection to SKOV3 Cells with Protamine-DOTAP/Chol Liposome-Oligodeoxynuleotides Complex
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Abstract:
Objective: To observe the transfer efficiency of protamine-DOTAP/Chol liposome-oligodeoxynuleotides complex to SKOV3 cells. Methods: Synthesized NF-κB decoy ODNs or scrambled ODNs labeled by FITC on the 5′ends was mixed with rationic DOTAP/cholesteral liposome and protamine, making the charge ratio of positive:negative 4. SKOV3 cells were transfected by the complex(1 μmol/L) and the efficiency were measured by flow cytometry(FCM) and fluorescence meter. The direct effect of NF-κB decoy ODNs transfer on the growth of SKOV3 cell line in vitro was measured by MTT method. Results: The efficiency of LPD transferto SKOV3 cell line was 96.41%, significantly higher than that of naked DNA transfer which was 12.86%.The growth of SKOV3 cell line was not affected by 48h incubation after NF-κB decoy ODNs or scrambled ODNs transfection.Conclusion: LPD method is the one with high transfer efficiency not affected by NBS(newborn bovine serum)with possibility of use in viv.
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Clc Number:
R730.3
