Lentivirus Packaging Kit
Based on 1 Customer Validation
MCE Lentivirus Packaging Kit is based on the second-generation lentiviral packaging system and is also compatible with both second- and third-generation packaging plasmids. Lentiviruses produced using this kit exhibit high infection efficiency and a broad host range, enabling stable and sustained expression of exogenous genes in host cells.
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Storage :
-20°C, 1 year, 4°C, 6 months. Avoid repeated freeze-thaw cycles.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
The lentiviral vector is a gene therapy vehicle derived from HIV-1, capable of stably integrating exogenous genes into the host cell genome. It features a broad host range, enables efficient infection of both dividing and non-dividing cells, and ensures long-term and stable expression of transgenes. Therefore, it has become a powerful tool for gene delivery, widely used in gene overexpression, RNA interference, microRNA studies, and in vivo animal experiments across various cell lines.
MCE Lentivirus Packaging Kit is based on the second-generation lentiviral packaging system and is also compatible with both second- and third-generation packaging plasmids. Lentiviruses produced using this kit exhibit high infection efficiency and a broad host range, enabling stable and sustained expression of exogenous genes in host cells.
Protocol
1. Preparation of Packaging Cells
Seed HEK293T or other packaging cells before transfection so that cell density is approximately 70%-80% at transfection and the cells are in good condition.
2. Packaging Reaction
| Component | Reference amount per packaging reaction |
|---|---|
| Transfer/Control Plasmid | 4 μg |
| Lentiviral Packaging Plasmid Mix | 4 μL |
| Lentivirus Transfection Reagent | 40 μL |
1. Dilute the Transfer Plasmid or Control Plasmid together with Lentiviral Packaging Plasmid Mix in a suitable serum-free medium.
2. Dilute Lentivirus Transfection Reagent separately.
3. Add the transfection reagent to the plasmid mixture, mix gently, and incubate for approximately 15 min at room temperature.
4. Add the complexes evenly to the packaging-cell culture and continue incubation.
3. Viral Harvest
1. Collect the first viral supernatant approximately 48 h after transfection and add fresh complete medium.
2. Collect viral supernatant again at approximately 72 h and pool the two harvests.
3. Centrifuge to remove cell debris, then filter, concentrate, or determine viral titer as required.
4. Viral Infection
Add an appropriate amount of Polybrene during infection according to the target cell type. Optimize viral dose and MOI experimentally.
Note: All lentivirus transfection, harvest, infection, and waste-disposal procedures should be performed under BSL-2 biosafety conditions.
Storage
-20°C, 1 year, 4°C, 6 months.
Avoid repeated freeze-thaw cycles.
Attention
1. Avoid repeated freeze/thaw cycles of plasmids. It is recommended to aliquot plasmids according to experimental needs and store them at -20°C for long-term preservation to ensure plasmid purity and transfection efficiency.
2. When collecting crude viral supernatant, in addition to routine aliquoting, it is advisable to prepare several small-volume aliquots (20-100 μL). Store these together with the main viral stock at -80°C. Once the small aliquots are fully frozen, thaw one aliquot and perform a titer determination. This simulates the titer reduction caused by a single freeze/thaw cycle, providing a more realistic and representative viral titer value that better reflects the actual infection efficiency.
3. All lentivirus-related procedures—including transfection, viral harvest, infection, and waste disposal—must be conducted in a BSL-2 (biosafety level 2) biosafety cabinet to ensure safe handling and containment.
4. This product is for R&D use only, not for drug, household, or other uses.
5. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K2030-10 T | HY-K2030-20 T | HY-K2030-40 T |
|---|---|---|---|
| Lentiviral Packaging Plasmid Mix | 40 μL | 80 μL | 160 μL |
| Control Plasmid (1 μg/μL) | 10 μL | 20 μL | 40 μL |
| Lentivirus Transfection Reagent | 400 μL | 800 μL | 1600 μL |
| Polybrene (10 mg/mL) | 200 μL | 400 μL | 800 μL |
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