Millipore Sigma Vibrant Logo
Attention: We have moved. EMD Millipore products are no longer available for purchase on emdmillipore.com.Learn More

70191 pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen

70191
Purchase on Sigma-Aldrich

Overview

Replacement Information

Products

Catalog NumberPackaging Qty/Pack
70191-3 Plastic ampoule 20 rxn
70191-4 Plastic ampoule 40 rxn
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-min reaction using premixed reagents. Following a 5-min heat inactivation step, the treated insert is combined with the ready-to-use vector and ligated in an optimized 15-min 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 Number70191
Brand Family Novagen®
Features and benefits
  • No restriction enzymes or special primers
  • Compatible with any DNA polymerase
  • Independent of 3ʹ-dA addition or sequence context of DNA ends
  • Blue/white screening with seven different vectors, including pETBlue expression vectors
  • Simple protocol takes less than one hour* from PCR product to plating transformants

* All times listed refer to kits using NovaBlue Singles™ Competent Cells and ampicillin or chloramphenicol selection. Giga kits, which use NovaBlue GigaSingles™ Competent Cells, require one hour for outgrowth.

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.

Product Information
Components
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
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
70191-3 04055977258486
70191-4 04055977258493

Documentation

pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen Certificates of Analysis

TitleLot Number
70191

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.

Brochure

Title
Express yourself
High fidelity gene amplification
The Complete Molecular Biology Toolkit - Expert workflow solutions from DNA cloning to protein expression

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

    Related Products & Applications

    Related Products

    Catalog Number Description
    70174 pT7Blue Blunt Vector (linearized) - Novagen
    70183 Introductory pT7Blue Perfectly Blunt® Cloning Kit - Novagen
    70184 Introductory pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen
    70188 pSTBlue-1 Blunt Vector (linearized) - Novagen
    70189 pT7Blue Perfectly Blunt® Cloning Kit - Novagen
    70633 Introductory pETBlue-1 Perfectly Blunt® Cloning Kit - Novagen
    70634 pETBlue-1 Perfectly Blunt® Cloning Kit - Novagen
    70635 Introductory pETBlue-2 Perfectly Blunt® Cloning Kit - Novagen
    70636 pETBlue-2 Perfectly Blunt® Cloning Kit - Novagen
    71229 pSTBlue-1 Perfectly Blunt® Giga Cloning Kit - Novagen

    Related Products

    Catalog Number Description
    70184 Introductory pSTBlue-1 Perfectly Blunt® Cloning Kit - Novagen
    70188 pSTBlue-1 Blunt Vector (linearized) - Novagen
    71229 pSTBlue-1 Perfectly Blunt® Giga Cloning Kit - Novagen

    Related Products By: Application Facete

    Related Products By: Brand Facete

    Categories

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