70598 pETBlue-1 AccepTor™ Vector Kit - Novagen

70598
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      Plastic ampoule 20 rxn
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      70598-4CN
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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.


          In addition to the pETBlue™ Systems, which contain uncut plasmids, pETBlue vectors are available in formats ready for insertion of PCR products. Two types of kits are available:
          AccepTor™ Vector Kits and Perfectly Blunt® Cloning Kits. The AccepTor Vector Kit enables direct insertion of DNA containing single 3′-dA overhangs, such as those amplified with non-proofreading DNA polymerases (e.g., NovaTaq™ DNA Polymerase), while the Perfectly Blunt Cloning Kits are designed for cloning inserts having any type of DNA end.






          Commercial use of this Product requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
          Catalogue Number70598
          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
          20 rxn40 rxn
          2 × 0.5 µg4 × 0.5 µgpETBlue-1 AccepTor Vector
          10 µl10 µlAccepTor Vector Positive Control Insert
          2 × 55 µl4 × 55 µlClonables 2X Ligation Mix
          1.5 ml1.5 mlNuclease-free Water
          22 × 50 µl44 × 50 µlNovaBlue Singles Competent Cells
          5 × 2 ml9 × 2 mlSOC Medium
          10 µl10 µlTest Plasmid
          2 × 0.2 ml4 × 0.2 mlTuner™(DE3)pLacI Competent Cells
          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
          Transport Information
          Supplemental Information
          Specifications

          Documentation

          pETBlue-1 AccepTor™ Vector Kit - Novagen Certificates of Analysis

          TitleLot Number
          70598

          Brochure

          Title
          The Complete Molecular Biology Toolkit - Expert workflow solutions from DNA cloning to protein expression

          Citations

          Title
        • Alla Korepanova, et al. (2005) Cloning and expression of multiple integral membrane proteins from Mycobacterium tuberculosis in Escherichia coli. Protein Science 14, 148-158.
        • Deanne M. Compaan and W. Ross Ellington. (2003) Functional consequences of a gene duplication and fusion event in an arginine kinase. 206, 1545-1556.
        • Paul L. DeAngelis and Amy J. Padgett-McCue. (2000) Identification and molecular cloning of a chondroitin synthase from Pasteurella multocida type F. Journal of Biological Chemistry 275, 24124-24129.
        • User Protocols

          Title
          TB248 AccepTor™ Vector Kits

          Vector Map

          Title
          TB258VM pETBlue™-1 Vector Map

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          Categories

          Life Science Research > Genomic Analysis > Transfection and Protein Expression > Bacterial Expression > Bacterial Expression Vectors
          Life Science Research > Genomic Analysis > DNA Preparation & Cloning > Cloning > Cloning Kits