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Invitrogen™ pLenti6/V5 Directional TOPO™ Cloning Kit

Product Code. 10562693
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Product Code. 10562693 Supplier Invitrogen™ Supplier No. K495510

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Description

Directional TOPO Cloning site for rapid and efficient directional cloning of blunt-end PCR products

  • Contains TOPO-adapted ViraPower lentiviral expression vector, pLenti6/V5-D-TOPO for quick PCR-based cloning and expression of a target gene in dividing and nondividing mammalian cells
  • Vector has CMV promoter for driving high-level, constitutive expression of the target gene and blasticidin selection marker for stable selection in mammalian cells
  • High efficiency and rapid cloning
  • Produces replication-incompetent virus for enhanced biosafety of system
  • Rouse Sarcoma Virus (RSV) enhancer/promoter for Tat-independent production of viral mRNA in the producer cell line
  • Modified HIV-1 5' and 3' Long Terminal Repeats (LTR) for viral packaging and reverse transcription
  • HIV-1 psi (ψ) packaging sequence for viral packaging
  • HIV Rev response element (RRE) for Rev-dependent nuclear export of unspliced viral mRNA
  • (CMV) immediate early promoter for high-level constitutive expression of gene of interest in mammalian cells
  • C-terminal V5 epitope for detection of recombinant protein
  • Blasticidin (bsd) resistance gene for selection in E. coli and mammalian cells
  • Ampicillin resistance gene for selection in E. coli
  • pUC origin for high-copy replication and maintenance of plasmid in E. coli

Constitutive Expression, Mammalian Expression, Protein Expression, Proteins, Expression, Isolation and Analysis, Viral Gene Delivery

TRUSTED_SUSTAINABILITY

Specifications

Cloning Method Directional TOPO™
Constitutive or Inducible System Constitutive
Content And Storage pLenti⁄V5-D-TOPO™ reagents (Box 1) Store at -20°C
• pLenti6⁄V5-D-TOPO™ Reagents (15-20 ng⁄μl linearized plasmid DNA in: 50% glycerol, 50 mM Tris-HCl, pH 7.4 (at 25°C),1 mM EDTA, 2 mM DTT, 0.1% Triton X-100, 100 μg⁄ml BSA, 30 μM bromophenol blue)
• dNTP Mix (12.5 mM each dATP, dCTP, dGTP, dTTP in water, pH8 at a total volume of 10 μl)
• Salt Solution (1.2 M NaCl and 0.06 M MgCl2, volume of 50 μl)
• Water (1 ml)
• CMV Forward Sequencing Primer (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 20 μl)
• Control PCR Primers (0.1 μg⁄μl each in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• Control PCR Template (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• pLenti6⁄V5-GW⁄lacZ (lyophilized in TE Buffer, pH 8, 10 μg)

One Shot™ Stbl3™ Chemically Competent E. coli (Box 2) Store at -80°C
• S.O.C. Medium (2% Trypton, 0.5% Yeast Extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2, 10 mM MgSO4, 20 mM glucose. 6ml) May be stored at +4°C or room temperature
• Stbl3™ Cells (21 x 50 μl)
• pUC19 Control DNA (10 pg⁄μl in 5 mM Tris-HCl, 0.5 mM EDTA, pH 8. 50 μl)
Delivery Type Lentiviral
For Use With (Application) Viral Expression
No. of Reactions 20 reactions
Product Line TOPO, ViraPower
Product Type TOPO Cloning Kit
Promoter CMV
Protein Tag V5 Epitope Tag
Quantity 20 Reactions
Selection Agent (Eukaryotic) Blasticidin
Vector pLenti, Directional TOPO Vectors
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For Research Use Only. Not for use in diagnostic procedures.

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