JRD352 stabilized cell line construction technology services
stable cell line construction technology Services
Stable cell line construction technology service service:
The basic principle of constructing a stable cell line is to clone the exogenous DNA to a carrier having a certain resistance, the carrier is transfected. To the host cell and integrated into the host chromosome, screening is performed using the resistance marks contained in the carrier. A commonly used eukaryotic expression vector resistance screening marker has neomromycin, hygromycin and puromycin, often used G418 instead of neomycin for selective screening, screening is stable Expressing the protein, or stabilizing the cell line of the silent specific gene.
The construction period of the stable cell line is long, difficult, each step is critical, especially cell transfection, common physical methods (electric rotation), chemical methods (liposome, etc.), biological method (virus), according to different Experimental conditions, choose a good method to achieve the purpose of experiment.
Shanghai AY
In the construction of stable cell lines, there is a wealth of experience, familiar with each step, especially for key steps, success Many scientific research customers construct a global cell line (overexpression, induced exposure, silence, induce silence, reporting gene, etc.). A, protein over expression Overgraduation of the protein is often a conventional means of studying the function of the gene / protein, and is also a method of enriched protein (DHFR, GS System) by means of expression systems. The construction of proteins over expressed stable cell lines can help to study this protein or give a large amount of protein. Shanghai's protein over-expressing stabilized cell line construction services construct a plasmid / virus containing the purpose of gene CDS by transfection / infection, and infection with the eukaryotic expression vector / lentiviral system. Using antibiotic screening, high copy clones with stable expression were obtained, and the construction of stable cell line was completed through 32 generation stability QC detection.
The following steps are mainly included:
1. Carrier construction (expression plasmid vector, viral vector);
2. Transfection / infection;
3. Purpose protein preliminary test;
4. Antibiotic Screening;
5. Purpose protein again (qualitative / semi quantitative);
6. Stability of monoclonal cell lines.
Special overter protein / cells, please consult in advance.
b, protein silence u0026Nbsp;
Knock-Down or Knock-out is often a conventional means of studying the gene / protein function, and the detection of cell phenotype after protein silencing is the gold standard method of research.
Shanghai AY
The protein silent stabilized cell line construction service constructs a gene that contains silent use of the silent expression carrier / slow viral system (shRNA) / microRNA) plasmid / virus, through transfection / infection, integrated silence genes on various cellular genome, using antibiotics, resulting in highly silent clones, after 32 generation stability QC detection, the construction of stable cell lines is completed. mainly includes the following steps:
1. Carrier construction (silence plasmid vector, viral vector); 2. Transfection / infection; 3. Purpose protein preliminary test; 4. Antibiotic Screening; 5. Purpose protein again (qualitative / semi quantitative);
6. Stability of monoclonal cell lines.
Special overter protein / cells, please consult in advance.
C, induced
Exogenous protein induced expression / silent can compare the difference between the bottom and intervention, very powerful interpretation of the mechanism of action of gene / protein, especially suitable for the study of toxic proteins It has been widely used.
Shanghai AY
Provides inducible protein expression / silent stable cell line construction services to construct u0026 ldquo; Switch u0026 rdquo; plasmid / virus controlled expression / silent genes, by transfection / infection, integrated carrier on various cellular genome, using antibiotic screening, resulting in induced stable cell line, after 32 generation stability Qc detection, completed Construction of stable cell lines. Mainly included with the following steps: 1. Carrier construction (induced plasmid vector, viral vector);
2. Transfection / infection; 3. u0026 Ldquo; switch u0026 rdquo; induction of protein preliminary detection 4. Antibiotic screening; 5. u0026 Ldquo; switch u0026 rdquo; purpose protein again detects (qualitative / semi-quantitative); 6. Stability of monoclonal cell lines.
Special overter protein / cells, please consult in advance.
1. The construction of stable cell lines is not the same, the difference is very large, the success rate is also very large, and the attitude towards the customer, it is recommended to consult in advance, and arrange the pre-arrangement Experiment;
2. Customers can provide gene sequences, carriers, cells, can also find the company's template library, carrier library, and cell library, can also beOur company purchasing;
3. This service only affects the expression, stability of the purpose, does not involve gene function, specific gene function detection, require additional evaluation and charge;
Submitted products and materials:
1. Constructing a good plasmid / virus (1 mL), contains a destination gene sequence / silencing sequence / induction sequence;
2. Tube 2 tube;
3. Experiment report.
At present, AY Pharma, Main Experimental Technical Services, Services, Animal Model Construction, Cell Straopers, ELISA Kits, etc. Welcome major scientific research enthusiasts to come to consult!