pOptiVEC™-TOPO™ TA Cloning™ Kit

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The OptiCHO™ Express Kit is designed for efficient growth and transfection of dihydrofolate reductase- deficient (DHFR-) Chinese hamster ovary (CHO) DG44 cells in suspension culture. The kit provides a complete workflow protocol as well as the necessary components for efficient transfection of DHFR- DG44 cells in a serum-free environment and subsequent stable cell line development, including:
• pOptiVEC™-TOPO® vector - a bicistronic cloning vector containing a DHFR gene for stable cell line development in CHO-DG44 cells (Figure 1); the vector is TOPO® adapted and enables cloning of a PCR product containing your gene of interest with >85% efficiency
• FreeStyle™ MAX Transfection Reagent - an animal origin-free transfection reagent offering high transfection efficiencies and consistent results
• CD OptiCHO™ Medium - a chemically defined, protein-free medium (without hypoxanthine or thymidine) specifically designed for high-density suspension cell culture of recombinant CHO-DG44 cells and secreted protein yield
• CD DG44 Medium - a chemically defined, protein-free medium supplemented with hypoxanthine and thymidine to support optimal growth of DG44 cells in suspension culture
• DG44 cells - preadapted to CD DG44 medium and selected for superior cell growth and transfection efficiencies

In addition, the OptiCHO™ Express Kit provides GIBCO® OptiPRO™ SFM for improved DNA-lipid complex formation, L-glutamine (provided separately for increased media stability), and Pluronic® F-68 to control shear forces in suspension cultures.

Store Pluronic® F-68 at 15°C to 30°C. Store CD DG44 Medium, OptiPRO™ SFM, CD OptiCHO™ Medium, and FreeStyle™ MAX Transfection Reagent at 2°C to 8°C in the dark. Store L-glutamine, pOptiVEC™-TOPO® vector, CMV Forward Sequencing Primer, and EMCV IRES Reverse Sequencing Primer at -5°C to -20°C. Store One Shot® TOP10 Chemically Competent E. coli cells at -80°C. Store DG44 cells in liquid nitrogen.
For Research Use Only. Not for use in diagnostic procedures.


Promoter: CMV
Key Function: Protein Production, Stable Cell Line Development
Protein Tag or Fusion: Untagged
Delivery Method: Transfection
Cloning Method: TOPO®-TA
Selection Agent (Eukaryotic): HT (-) Media, MTX (Methotrexate)
Vector Type: TOPO-TA Vectors
Constitutive or Inducible System: Constitutive
Product Line: Gibco®
Product Size: 1 kit
Special Elements: EMCV IRES


Manuals & protocols

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