规格
Cloning Method:	TOPO®-TA
Key Functions:	Constitutive Expression
Product Line:	TOPO®, pcDNA™
Constitutive or Inducible System:	Constitutive
Delivery Type:	Transfection
Promoter:	CMV
Selection Agent (Eukaryotic):	Geneticin® (G-418)
Quantity:	40 reactions
Vector:	TOPO-TA Vectors, pcDNA
Protein Tag:	His Tag (6x), V5 Epitope Tag

储存

2 × 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)

2 × 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

描述

The pcDNA™3.1/V5-His TOPO® TA Expression Kit offers one-step cloning of Taq-amplified PCR products into a high-level expression vector. Topoisomerase activation of the pcDNA3.1/V5-His-TOPO® vector allows PCR products to be ligated in just 5 minutes on your bench top and results in 90% recombinants.

In addition, the vector includes 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.

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