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Summary of virus packaging experiment experience
First, the effects of cell status , vector system, and gene of interest on viral packaging (gene size, sequence, protein functional toxicity, etc. ) are three major factors affecting virus quality.
Second, the cell state needs to be judged by rich cell culture experience . For example, before packaging or transfection, it is necessary to pay attention to whether the cells are clean and fine polluted, whether the fullness is good, and the uniform cell density is moderate . Also when transfected cell density must pay great attention, usually in the 70% or so.
Third, it is recommended to use a commercialized mature carrier system .
Fourth, as for the operational control details and transfection reagent factors during packaging transfection, it is necessary to pay attention to the fact that the cells are not contaminated in the middle of the process and the entire process needs to be repeated .
Fifth, another concern is the expression of the gene construct and the extraction and purification of the links, the purified extraction for contamination, has reached the purity requirements. These links are also the key to ensuring the quality of the virus .
Sixth, the size of the target gene sequence, the function of the toxicity of the protein of interest such as lead can affect the success of the package or not, we can replace the viral vector packaging system to try another carrier systems to achieve the purpose.
Virus Packaging Technology 8 - Summary of Virus Packaging Experiment Experience
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