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70597 Introductory pETBlue-1 AccepTor™ Vector Kit - Novagen

70597
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Overview

Replacement Information

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Catalog NumberPackaging Qty/Pack
70597-3 Plastic ampoule 10 rxn
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.

Catalogue Number70597
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
0.5 µgpETBlue-1 AccepTor Vector
10 µlAccepTor Vector Positive Control Insert
55 µlClonables 2X Ligation Mix
1.5 mlNuclease-free Water
11 × 50 µlNovaBlue Singles Competent Cells
3 × 2 mlSOC Medium
10 µlTest Plasmid
0.2 mlTuner™(DE3)pLacI Competent Cells
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
Transport Information
Supplemental Information
Specifications

Documentation

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

TitleLot Number
70597

Brochure

Title
High fidelity gene amplification

Citations

Title
  • 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
    TB053 Academic and Non-profit Laboratory Assurance Letter
    TB248 AccepTor™ Vector Kits

    Vector Map

    Title
    TB258VM pETBlue™-1 Vector Map

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    70598 pETBlue-1 AccepTor™ Vector Kit - Novagen
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    71228 pSTBlue-1 AccepTor™ Vector Giga Kit - Novagen

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    Categories

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