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70594 Introductory pSTBlue-1 AccepTor™ Vector Kit - Novagen

70594
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      Description
      Overview

      The AccepTor™ Vector Cloning Kits are designed to simplify the PCR cloning process by using non-proofreading thermostable DNA polymerases (i.e. KOD XL polymerase and native and recombinant Taq polymerases), which leave single 3'-dA overhangs on the reaction products. The AccepTor Vectors enable direct ligation by virtue of single 3′-dU overhangs that anneal efficiently with 3′-dA extensions on PCR products. The dU residues are converted to dT residues in vivo following transformation.

      AccepTor Vectors are supplied in a ready-to-ligate format. Simply mix the vector with your PCR product, add Clonables™ 2X Ligation Premix (Cat No. 70573), and transform into NovaBlue Singles™ Competent Cells (Cat. No. 70181). The entire procedure, from PCR product to plating transformants, can be performed in as little as 40 minutes with minimal pipetting.

      AccepTor Vector Kits are available in an introductory 10-reaction size as well as 20- and 40-reaction configurations. The AccepTor Vectors are also available separately without the ligation and transformation components. Also see pSTBlue-1 AccepTor Vector Giga Kit (Cat. No. 71228) for information on using higher-efficiency competent cells.



      Two different vectors, pSTBlue-1 and pETBlue™-1, are available as AccepTor™ Vector kits. Each is carefully prepared and tested for optimal cloning efficiency and provides easy visualization of recombinants by blue/white screening using lacZ α-complementation. The pSTBlue-1 is a general purpose vector with dual opposed T7 and SP6 promoters, both with amp and kan resistance, and an array of flanking restriction sites. The pETBlue-1 vector is a novel plasmid specifically designed to enable high-level T7 RNA polymerase-driven expression of target genes, while providing the convenience of a blue/white screening method. Screening is independent of expression because the associated T7lac expression promoter is in an opposed orientation relative to the E. coli promoter. pETBlue-1 facilitates the expression of native unfused proteins and allows for convenient subcloning of target genes already fused to existing detection and purification tags. An EcoR V cloning site is appropriately spaced downstream of an E. coli ribosome binding site. Inserts must encode an ATG start codon at their 5′ end if expression is desired. The sequence is numbered from the first base of the T7 promoter sequence.

      Catalogue Number70594
      Brand Family Novagen®
      Features and benefits• No restriction enzymes or special primers
      • Direct ligation of PCR product with vector
      • Compatible with polymerases that leave single 3´-dA overhangs
      • Blue/white screening with pSTBlue-1 or pETBlue™-1 vectors
      • Simple protocol takes as little as 40 minutes from PCR product to plating transformants
      References
      Product Information
      Components
      DeclarationThis product is covered by U.S. Patent 5,856,144 owned by EMD Chemicals Inc. or its Affiliates.
      Quality LevelMQ100
      Applications
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage ≤ -70°C
      Do not freeze Ok to freeze
      Packaging Information
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      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      70594-3 04055977273267

      Documentation

      Introductory pSTBlue-1 AccepTor™ Vector Kit - Novagen Certificates of Analysis

      TitleLot Number
      70594

      Brochure

      Title
      High fidelity gene amplification

      Citations

      Title
    • Tsui-Fen Chou, et al. (2005) 31P NMR and genetic analysis establish hinT as the only E. coli purine nucleoside phosphoramidase and as essential for growth under high salt conditions. Journal of Biological Chemistry 15356-15361.
    • Anna B. Gilg, et al. (2005) Isolation and functional expression of an animal geranyl diphosphate synthase and its role in bark beetle pheromone biosynthesis. Proceedings of the National Academy of Sciences (USA) 102, 9760-9765.
    • James E. Turner, et al. (2005) Characterization of Arabidopsis fluoroacetate-resistant mutants reveals the principal mechanism of acetate activation for entry into the glyoxylate cycle. Journal of Biological Chemistry 280, 2780-2787.
    • Ruben Vicente, et al. (2005) Pattern of Kvb subunit expression in macrophages depends upon proliferation and the mode of activation. Journal of Immunology 174, 4736-4744.
    • User Protocols

      Title
      TB248 AccepTor™ Vector Kits

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      Categories

      Life Science Research > Genomic Analysis > DNA Preparation & Cloning > Cloning > Cloning Kits