产品介绍
Product Description
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规格 |
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| 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: | 20 reactions |
| Vector: | Directional TOPO Vectors, pcDNA |
| Protein Tag: | His Tag (6x), V5 Epitope Tag |
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储存 |
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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) 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 |
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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.
