规格
Cloning Method:	Directional TOPO®
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:	Directional TOPO Vectors, pcDNA
Protein Tag:	His Tag (6x), V5 Epitope Tag

储存

2 × Box 1 (Store at -20°C):
• 20 µl pcDNA™3.1D/V5-His-TOPO® (15–20 ng/µl)
• 10 µl dNTP Mix
• 50 µl Salt Solution
• 1 ml Sterile Water
• 20 µl T7 Sequencing Primer (0.1 µg/µl)
• 20 µl BGH Reverse Sequencing Primer (0.1 µg/µl)
• 10 µl Control PCR Template (0.1 µg/µl)
• 10 µl Control PCR Primers (0.1 µg/µl each)
• 10 µl Expression Plasmid (0.5 µg/µl)

2 × Box 2 (Store at -80°C):
• 6 ml S.O.C. Medium (may be stored at 4°C or room temperature)
• 21 × 50 µl TOP10 Cells
• 50 µl pUC19 Control DNA

描述

The pcDNA™3.1 Directional TOPO® Expression Kit uses linearized, topoisomerase I-activated pcDNA3.1D/V5-His-TOPO® for five-minute directional cloning and subsequent high-level expression.

Directional Cloning technology facilitates expression experiments because:

• A proofreading enzyme is used for fewer errors in cloned genes
• Greater than 90% of the clones are in the correct orientation for expression, reducing time spent colony screening for clone orientation

In addition, pcDNA3.1D/V5-His-TOPO® provides strong expression levels from the CMV promoter and the option of a C-terminal V5-His fusion tag for easy detection of recombinant protein with an Anti-V5 Antibody and rapid purification on nickel-chelating resin.

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