JRD1000 slow virus packaging technology detection service

Product introduction : Lentivirus carrier is HIV-1 (human immunodeficiency type 1 virus) The gene therapy vector developed by the foundation. It has a wide infection spectrum, which can effectively infect splitting period and static cells, and long-term stable expression of exogenous genes, so as a powerful tool for introducing foreign genes. The slow virus system is now widely used to gene overcaries, RNA interference, microRNA study, and living animal experiments. Slow Virus Packaging Technology Chronic Virus Packaging Technology Testing Service

Company provides a slow viral vector construction, RNAI (shRNA), miRNA, overexpression and interference lentiviral packaging, and stable cell line construction services based on slow virus technology.

company's slow virus products are the fourth-generation plasmid system, which features high expression, high titer, rapid cloning and Easy to test. Slow virus titers can reach 1e + 8 Tu / ml, without concentration and purification, infect the purpose of cells and screening through GFP, the experimental requirements of different experiments can make subsequent experiments more convenient and fast.

(1) Experimental process (1, 2 3 is a side steps): 1. Construction of a lentivirus overgraduated plasmid vector to create a top-down specific amplification primer while introducing an enzyme dug The PCR technology (using high-fidelity KOD enzymes, 3K mutation rate is 0%) from the template (cDNA plasmid or library) to facilitate the T-vector Coding Sequence. The CDS region was cut from the T support, and the transacted vesicle was mounted.

2. Constructing a Sribunic SiRNA-cervical ring structure of a synthetic siRNA in a lentiviral interfering massage medium, annealing, a slow viral interfering massage carrier.

3. MiRNA vector constructs a MIR precursor that is adjusted from the Genomic and introduces an enzyme dug in the tub.

4. Packaging and concentration purification of slow viral vectors and its auxiliary packaging original carrier plasmid (two plasmid packaging system), three plasmid vectorsThe high purity non-tank extraction was performed, and 293T cells were transfected. After transfection, the transfection was replaced with medium, cultured 48 and 72 h, and the cell supernatant rich in a slow viral particles was collected, and the viral supernatant passedUltra-centrifidal concentration virus.