70184 Introductory pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen

70184
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      Description
      Overview

      The Perfectly Blunt® Cloning Kits are designed for simplified cloning of DNA generated by PCR using any type of DNA polymerase. This approach enables the use of high-fidelity proofreading enzymes for amplification, thus decreasing the probability of generating mutations in the target sequence. In addition, under many conditions blunt cloning is more efficient than T-cloning, most likely due to the observation that the efficiency of single dA addition by Taq DNA polymerase varies significantly depending on the sequence context of the DNA ends, and even the number of PCR cycles performed (Novy 1996, Clark 1998, Brownstein 1996, Magnuson, 1996, Hu 1993).

      With the Perfectly Blunt cloning protocol, you can go from PCR product to plating transformants in less than one hour with minimal hands-on time. The finished PCR product is converted to a blunt, phosphorylated form in a 15-minute reaction using premixed reagents. Following a 5-minute heat inactivation step, the treated insert is combined with the ready-to-use vector and ligated in an optimized 15-minute reaction. An exclusive 8-minute transformation procedure using highly efficient NovaBlue Singles™ Competent Cells (Cat. No. 70181) generates recombinant colonies that are easily visualized by blue/white screening.

      Note that the Perfectly Blunt method is not limited to cloning PCR products; these kits are also suitable for cloning restriction fragments, cDNA, or sheared DNA with the same protocols.

      Seven different vectors are available in Perfectly Blunt® Cloning Kits. Vector choices include those designed for general cloning, sequencing, optimal in vitro transcription/translation, and optimal protein expression in E. coli. Each vector is available in a kit containing sufficient reagents for 10, 20, or 40 reactions.

      “Vector only” kits are also available in 20- and 40-reaction sizes without ligase and competent cells. For higher efficiency competent cells, also consider pSTBlue-1 Perfectly Blunt® Giga Cloning Kit (Cat. No. 71229).

      The pSTBlue-1 vector is a multi-purpose cloning vector featuring a versatile multiple cloning region, blue/white screening, dual opposed T7/SP6 promoters and dual kanamycin/ampicillin resistance. Restriction sites producing 4-base 3′ overhangs are conveniently positioned near each end of the polylinker to facilitate the generation of unidirectional deletions using exonuclease III.



      Catalogue Number70184
      Brand Family Novagen®
      References
      References

      Novy, R.E., et al. 1996. inNovations 6, 7. Clark, J.M. 1988. Nucleic Acids Res. 16, 9677. Brownstein, J.M., et al. 1996. BioTechniques 20, 1004. Magnuson, V.L., et al. 1996. BioTechniques 21, 700. Hu, G. 1993. DNA and Cell Biology 12, 763.

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      Quality LevelMQ100
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      Ship Code Dry Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage ≤ -70°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
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      Introductory pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen Certificates of Analysis

      TitleLot Number
      70184

      References

      Reference overview

      Novy, R.E., et al. 1996. inNovations 6, 7. Clark, J.M. 1988. Nucleic Acids Res. 16, 9677. Brownstein, J.M., et al. 1996. BioTechniques 20, 1004. Magnuson, V.L., et al. 1996. BioTechniques 21, 700. Hu, G. 1993. DNA and Cell Biology 12, 763.

      Citations

      Title
    • Thomas S. Lendvay, et al. (2007) Compensatory paracrine mechanisms that define the urothelial response to injury in partial bladder outlet obstruction. American Journal of Physiology: Renal 293, F1147-F1156.
    • Neelima Sukumar, et al. (2007) Differential Bvg phase-dependent regulation and combinatorial role in pathogenesis of two Bordetella paralogs, BipA and BcfA. Journal of Bacteriology 189, 3695-3704.
    • Korry J. Hintze and Elizabeth C. Theil. (2005) DNA and mRNA elements with complementary responses to hemin, antioxidant inducers, and iron control ferritin-L expression. Proceedings of the National Academy of Sciences (USA) 102, 15048-15052.
    • Margo E. Mancl, et al. (2005) Two discrete promoters regulate the alternative-spliced human interferon regulatory factor-5 isoforms: multiple isoforms with distinct cell type-specific expression, localization, regulation and function. Journal of Biological Chemistry 280, 21078-21090.
    • M. Shimizu, et al. (2005) Salmon serum 22 kDa insulin-like growth factor-binding protein (IGFBP) is IGFBP-1. Journal of Endocrinology 184, 267-276.
    • Eric A. Grovender, et al. (2004) Single-chain antibody fragment-based Adsorbent for the extracorporeal removal of β2-microglobulin. Kidney International 65, 310-322.
    • User Protocols

      Title
      TB183 Perfectly Blunt® Cloning Kits

      Vector Map

      Title
      TB214VM pSTBlue-1 Vector

      Vector Sequence

      Title
      pSTBlue-1 Sequence

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      Categories

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