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Invitrogen™

pcDNA™3.3-TOPO™ TA Cloning™ Kit

Catalog number:  K830001

Related applications:

Antibody & Cell Culture Production

| Protein Expression

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Description

The pcDNA™3.3-TOPO® TA Cloning® Kit is designed to deliver exceptionally high levels of transgene expression in adherent cells as well as in the FreeStyle™ MAX CHO and FreeStyle™ MAX 293 suspension cell systems. The pcDNA3.3™-TOPO® TA Cloning® Kit is supplied with the TOPO®-adapted plasmid vector, all reagents for cloning, and One Shot® TOP10 competent cells. The pcDNA™3.3-TOPO® vector offers the following benefits:

• TOPO® adapted for cloning PCR products with >85% efficiency
• High-level gene expression in a wide range of mammalian cells due to the native cytomegalovirus (CMV) immediate-early promoter/enhancer sequence (672 bp)
• Vector is ideal for large-scale protein expression
• Generation of native protein without extraneous amino acids

The pcDNA™3.3-TOPO® vector is also included with the OptiCHO™ Antibody Express System for cloning and expression of light and heavy antibody chains.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Cloning Method: TOPO®-TA
Key Functions: Protein Production, Constitutive Expression
Product Line: TOPO®, pcDNA™
Constitutive or Inducible System: Constitutive
Delivery Type: Transfection
Promoter: CMV
Selection Agent (Eukaryotic): Geneticin® (G-418)
Vector: pcDNA
Protein Tag: Untagged

Contents & storage

Box 1 (store at -20°C)
• 20 µl pcDNA™3.3-TOPO® vector, TOPO® adapted (5–10 ng/µl)
• 100 µl 10X PCR Buffer
• 10 µl dNTP Mix
• 50 µl Salt Solution
• 1 ml Sterile Water
• 10 µl Control PCR Template (50 ng/µl)
• 10 µl Control PCR Primers (100 ng/µl each)
• 20 µl CMV Forward Sequencing Primer (100 ng/µl)
• 20 µl TK polyA Reverse Sequencing Primer (100 ng/µl)
• 10 µl pcDNA™3.3-TOPO®/lacZ Expression Control Plasmid (500 ng/µl)

Box 2 (store at -80°C)
• 11 × 50 µl TOP10 E. coli
• 50 µl pUC19 Control DNA (10 pg/µl)
• 6 ml S.O.C. Medium