RayBiotech's stable cell line service uses a mammalian retrovirus to incorporate foreign genes into the host genome. Our comprehensive service is customized to meet your needs (Table 1).
Table 1. The phases and descriptions of the custom stable cell line service
|Service Phase||Description**||Lead time||Deliverable|
|1: Gene Synthesis||The gene-of-interest is synthesized.||2 - 4 weeks||Gene will remain at RayBiotech for Phase 2. If desired, the gene can also be shipped to the customer.|
|2: Gene Cloning||The gene-of-interest will be cloned into a retrovirus plasmid. Proper insertion of the gene is confirmed via DNA sequencing.||2 weeks||Cloned gene will remain at RayBiotech for Phase 3.|
|3: Retroviral Packaging||The retrovirus is packaged using human GP2-293 cells. The retrovirus titer is determined using a plaque assay. Options for shipping include:
Mini-scale production (3-5 ml of virus; concentrated viral stock not available)
Standard production (200 µL concentrated viral stock; ∼1E8 to ∼1E9 pfu/ml)
Large-scale production (1 ml concentrated viral stock; ∼1E8 to ∼1E9 pfu/ml)
|2 - 3 weeks||
Images of the plaque assay and an Excel file with the associated data will be sent to the customer electronically.
The packaged retrovirus will remain at RayBiotech for Phase 4. If desired, the packaged retrovirus can be shipped to the customer.
|4: Stable Cell Line Generation||Cells are transduced with the gene-of-interest. Stable gene expression for 5 – 10 clones across 3 – 6 different passages will be confirmed with a western blot.||1 - 2 months||Western blot results will be sent electronically to customer. Stable cell line will remain at RayBiotech for Phase 5. If desired, the clone with the highest gene expression according to the western blot can be shipped to the customer.|
|5: Cell Culture & Treatment||RayBiotech can culture and treat the stable cells according to your specifications. Data analysis included.||Depends on experiment||Data will be sent electronically to customer. If desired, the stable cell line can be shipped to the customer.|
** Either RayBiotech or you can provide the materials.