This manual details the procedures for constructing lentivectors, packaging them in 293T cells, and infecting target cells, including virus concentration…
This manual provides a comprehensive guide for laboratory personnel on constructing lentivectors and packaging them using a three-plasmid system. It covers the preparation of shuttle plasmids and packaging vectors, followed by transfection into 293T packaging cells. The process includes detailed steps for cell culture, transfection using LipoFiter reagents, and the collection of viral supernatants. This manual is written in Chinese.
| Packaging Cell Line | 293T |
|---|---|
| Strain for Plasmid Amplification | DH5α |
| Transfection Reagent | LipoFiterTM |
| Plasmid Concentration Requirement | >1ug/ul |
| Polybrene Working Concentration | 6～8μg/ml |
The most common working concentration for Polybrene is 6～8μg/ml.
Collect the viral supernatant at 48 and 72 hours post-transfection.
The standard final concentration range for Puromycin is 1～10μg/ml.