2017•实验科学与技术Requires access
Experiment Teaching Reform of Cationic Liposome-mediated Gene Transfection
Liu Yanhong, Jing Li, Zhong Ji, Yu Xiaoqi, Kun Li
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Abstract
By optimizing the conditions of gene transfection,we explored the effect of the gene transfection conditions,such as 【liposome】/【DNA】 ratios,incubation time and exposure time of the complexes with cells,on gene transfection efficiency.Gel retardation assay was used to fix the 【liposome】/【DNA】 ratios,and then the effects of such combination ratios and times on the gene transfection efficiency were examined.Moreover,the quantity of DNA uptaked by cellular was further analyzed by using the confocal laser scanning microscope.According to the results,when the ratio of 【liposome】/【pEGFP】 is 1.25,gene transfection efficiency is the highest.After 4 hours of transfection,most of DNAs have been uptaked by the cells.It's time to replace fresh culture medium.