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pcDNA™3.1/V5-His TOPO® TA Expression Kit

Catalog number: K480001

Invitrogen™  Related applications: Protein Expression

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The pcDNA™ 3.1/V5-His TOPO® TA Expression Kit provides a highly efficient, 5-minute, one-step cloning strategy ("TOPO® Cloning") for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector. No ligase, post-PCR procedures, or PCR primers containing specific sequences are required. Once cloned, analyzed, and transfected, the PCR product will express directly in mammalian cell lines. In addition, the pcDNA™ 3.1/V5-His TOPO® vector (see figure) has the following features:

• Strong CMV promoter for high-level, constitutive expression
• C-terminal V5 epitope tag for efficient detection of recombinant proteins with an Anti-V5 antibody
• C-terminal polyhistidine (6xHis) sequence for purification using nickel-chelating resin and detection with an Anti-His (C-term)antibody
For Research Use Only. Not for use in diagnostic procedures.


Promoter: CMV
Key Function: Constitutive Expression
Protein Tag or Fusion: His Tag (6x), V5 Epitope Tag
Delivery Method: Transfection
Cloning Method: TOPO®-TA
Selection Agent (Eukaryotic): Geneticin® (G-418)
Vector Type: pcDNA, TOPO-TA Vectors
Product Size: 20 reactions
Constitutive or Inducible System: Constitutive

Contents & storage

Box 1 (store at -20°C)
• 20 µl pcDNA™3.1/V5-His-TOPO® (10 ng/µl)
• 100 µl 10X PCR Buffer
• 10 µl dNTP Mix (50 mM)
• 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 T7 Sequencing Primer (100 ng/µl)
• 20 µl BGH Reverse Sequencing Primer (100 ng/µl)
• 10 µl Expression Control Plasmid (500 ng/µl)

Box 2 (store at -80°C)
• 21 × 50 µl TOP10 E. coli cells
• 50 µl pUC19 Control DNA (10 pg/µl)
• 6 ml S.O.C. Medium


Manuals & protocols

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