* This product is for research use only. Not intended for use in the treatment or diagnosis of disease.
The classification of viral vector systems includes:
Depending on the intended application, unidirectional or bidirectional cDNA libraries can be generated. Complementary DNA is generated using oligo-DT primers or random hexamer primers.
When the virus is added to the target cell, the genetic material is shuttled into the cell where it is randomly integrated into the host genome. Any genes placed between the LTR or ITR during vector cloning are permanently inserted into the host's DNA along with the rest of the viral genome.
1. Construct viral plasmid vector
2. Transfect packaged cells
Transport plasmids, envelope plasmids and gag-pol plasmids were co-transfected with packaging cells (such as cells derived from HEK293T were highly transfected with different transfection reagents).
Some transfection reagents are available for the production of viral vectors:
Product Information | |
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Product Name | Broad Spectrum DNA Transfection Reagent |
Catalog Number | BT-000023 |
Shelf Life | 1 year. |
Storage | Store at 4°C. |
Cell Densitiy and Confluency | Ideally, cells should be ≥80% confluent prior to transfection. |
Characteristics | DNA transfection reagent for various cells. Low cytotoxicity. High transfection efficiency. |
Product Information | |
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Product Name | Lentiviral Transfection Reagent |
Catalog Number | BT-000028 |
Shelf Life | 6 months, when properly stored and handled. |
Storage | Store at 4°C. |
Cell Densitiy and Confluency | Optimally, ~90% confluency. |
Characteristics | High transfection efficiency in HEK293 cells. High titer lentivirus production. Serum compatibility, no need to change any medium during transfection. |
Product Information | |
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Product Name | Virus DNA Transfection Reagent |
Catalog Number | BT-000027 |
Shelf Life | 6 months, when properly stored and handled. |
Storage | Store at -20°C. |
Cell Densitiy and Confluency | Recommended 80-95% confluence at the time of transfection for adherent HEK 293T/17 cells, 2 x 106 cells/ml for suspension 293-F cells. |
Characteristics | Efficient DNA delivery. Flexible-compatible with different recombinant viruses and cell culture systems. No animal sources. Reliable-stable high virus titer production. Culture format versatility. |
Product Information | |
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Product Name | Large RNA Transfection Reagent |
Catalog Number | BT-000040 |
Shelf Life | 1 year. |
Storage | Store at 4°C. |
Cell Densitiy and Confluency | 60%-80% while transfection. |
Characteristics | Low cytotoxicity. High transfection efficiency -enable efficient RNA transfection in a variety of cell lines to ensure successful experiments. Serum compatible -transfection in the presence of serum without changing media. Suitable for all sizes of RNA -especially large RNA and CRISPR guide RNA. |
3. Collect the virus pellets
The time to reach maximum virus concentration is usually 48-72 hours after incubation. The culture medium was removed and filtered through a 0.45μm filter to obtain culture supernatant containing short-term production of virus, stored at -70°C or -80°C, or immediately used for infection of other packaged cell lines.
4. Purified virus
5. Virus titer
The common method is to infect the target cells with the virus, which can be roughly quantified according to the presence of vector RNA or protein.
References