The pLKO1 plasmid, pLenti entry-destination system,57 (link) and helper vectors, VSVG and delta8.2, were purchased from Addgene. Virus packaging and target cell infection were performed following standard protocols from Addgene. Stable cell lines were created by selection with 1 μg/ml puromycin (P9620, Sigma) for 6 days.
Protocol detail
pLKO1 Lentiviral Transduction Protocol
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Corresponding Organization :
Other organizations : Xiangya Hospital Central South University, Central South University, Harvard University
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Variable analysis
independent variables
- PLKO1 plasmid
- PLenti entry-destination system
- Helper vectors VSVG and delta8.2
- Stable cell lines
- Concentration of puromycin (1 μg/ml)
- Duration of puromycin selection (6 days)
- Not explicitly mentioned
- Not explicitly mentioned
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