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Some cells are exceptionally large or small, especially certain primary cells, and some cells aggregate, making counting difficult. The approach for these cells may vary from the preliminary experiments. During the pilot experiment, maintain a cell confluence of 30-40% at the time of infection. Even if the cells do not reach the predefined count of 10,000 cells/well, try to control the cell confluence around this quantity for the main experiment. For cells that form clusters, such as pancreatic islet cells or embryonic stem cells, pay attention to the number and size of cell clusters for futur…

Lentivirus may exhibit some toxicity towards your target cells. Adjust and reduce the MOI (Multiplicity of Infection) during infection. Additionally, consider changing the medium at 4 hours, 8 hours, and 12 hours post-infection with fresh complete culture medium for continued observation and cultivation.

The GFP fluorescence intensity in target cells depends on several factors, including the number of virus particles infecting the cells, the proliferation status of the cells, cell type, and the observation time. Generally, the more virus particles infect the target cells, and the faster the cells proliferate, the stronger the GFP fluorescence will be. Lentivirus is a slow virus, and typically, the peak expression of the GFP gene is reached 48-72 hours after virus infection in rapidly proliferating cells. For cells with slower proliferation rates, the expression time of the GFP gene may be exte…

Typically, we increase the MOI (Multiplicity of Infection) to enhance viral transduction efficiency. When necessary, Polybrene (4-10 µg/mL) can be added to the culture medium to increase the infection rate of the virus. Additionally, ensuring optimal growth conditions for the target cells is crucial for achieving a normal infection efficiency.

The recommended MOI (Multiplicity of Infection) for common cell types can vary and is influenced by factors such as cell growth status, the type of infecting virus, and experimental conditions. Generally, the recommended MOI range is between 0.5 to 5. However, the optimal MOI may need to be optimized based on specific experimental conditions.Here are some recommended MOI ranges for common cell types, but please note that these are only references, and it is advisable to adjust based on specific experimental conditions:1. **HEK293 cells:** 1-52. **HeLa cells:** 1-53. **THP-1 cells:** 5-104. **H…

When the MOI value required for cell infection reaches 80%, it is defined as the optimal MOI value for these cells.Polybrene: Commonly used infection additive at a concentration of 5~10 µg/mL. Polybrene is a positively charged small molecule that binds to the anions on the cell surface, significantly enhancing virus-cell contact and increasing virus infection efficiency. It can improve infection efficiency by 3~4 times for general cells and up to 10~20 times for certain cells.1.1 Experiment12-well plate, pipette gun, tips, EP tubes, Polybrene, HBSS, waste liquid1.2 Experiment1.2.1 Adherent Ce…

The lentiviral vector provided by Aigi belongs to the "second generation" lentiviral vector. The enhancer function of the 3' LTR in its genome is deleted, forming the so-called "self-inactivation" (SIN). After the viral genome integrates into the cell genome, it does not produce new progeny viruses, reducing the risk of unintended activation of surrounding genes, thus enhancing safety. However, the virus still carries potential biological risks, so it is recommended not to use pseudotyped viruses encoding known or potentially carcinogenic genes. Unless a gene is fully r…

1. Upon receipt, store the virus solution at 4°C (please use within one week). For long-term storage, keep it at -80°C. Avoid repeated freeze-thaw cycles as it may reduce virus titers; generally, viruses can be stored at -80°C for approximately six months. If stored for more than six months, reevaluate the virus titer before use.2. Lentivirus provided by Aigi is dissolved in HBSS (pH 7.4). When using the virus, thaw it in an ice bath after retrieving it from the -80°C freezer and store it at 4°C. If dilution is required, add an appropriate amount of HBSS for uniform mixing, or dilute it w…