Li WuFeng

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Jinsheng Wang

Changzhi Medical College

Jinbao Chen

Shanghai Pudong New Area Gongli Hospital

Chengping Zhao

Shanxi Agricultural University

XU Shang-zhong

Institute of Animal Sciences

Runjun Yang

The First Affiliated Hospital, Sun Yat-sen University

Lupei Zhang

Chinese Academy of Agricultural Sciences

Li Li

Sichuan Agricultural University

Gao Xue

Peking University Cancer Hospital

Junya Li

Chinese Academy of Sciences

Li Huifeng

Anhui University of Technology

Cooperative Institutions

Chinese Academy of Sciences

Sun Yat-sen University

Institute of Animal Sciences

Chinese Academy of Agricultural Sciences

Ministry of Agriculture and Rural Affairs

Kunming Medical University

Shanxi Agricultural University

The First Affiliated Hospital, Sun Yat-sen University

Zhongshan Hospital

Fudan University

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Optimization of transfection conditions for tranfecting pAcGFP-bFADD recombinant plasmid to Hela cell mediated by lipofectamine 2000.

Journal of Pharmaceutical and Biomedical Sciences (2009)

Runjun Yang Li WuFeng Lupei Zhang XU Shang-zhong Junya Li

pAcGFP-bFADD plasmid with AcGFP report gene was transfected to Hela cell mediated by lipofectamine 2000, and the transfection condition was optimized by adjusting transfection time and concentrations of plasmid DNA and liposome.Twelve hours after transfected, positive cells count was made under fluorescence microscope, and the cell viability was detected by MTT.Results indicated that perfect cell transfection efficiency(62.3% ) was obtained on the condition of the complex of 2.4 μg pAcGFP-bFADD and 3.0 μL lipofectamine 2000 transfecting Hela cell after 6 h, meanwhile, its cell viability could reach 83%.When the concentration of plasmid DNA and liposome increased to 3.2 μg and 6.0 μL respectively, the complex would bring higher toxicity, the inhibition rate reached 42.98%, the difference was extremely significant(P 0.01) compared with control group.After condition optimization, AcGFP-bFADD fusion gene got the expression peak 72 h after transfection.The green fluorescent positive cells ratio could reach 64%.

Lipofectamine

HeLa

Viability assay

MTT assay

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Establishment of the plasmid standard curve generation method for absolute quantification PCR.

Journal of Pharmaceutical and Biomedical Sciences (2011)

Li Li Chengping Zhao Hong Li Li WuFeng Lihuan Zhang

Standard curve

Absolute (philosophy)

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