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Merck - 70191-3 - pSTBlue-1 Perfectly Blunt Cloning Kit - Novagen

  • MFR SKU: 70191-3
  • ITEM #: 3441165
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ITEM #: 3441165
pSTBLUE-1 BLUNT KIT
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Merck - 70191-3 - pSTBlue-1 Perfectly Blunt Cloning Kit - Novagen

  • Item #: 3441165
  • MFR SKU: 70191-3
Technical Specs
  • 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 Nova. Blue 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 Number 70191
  • Brand Family Novagen®
  • Features and benefits
  • No restriction enzymes or special primers
  • Compatible with any DNA polymerase
  • Independent of 3-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 Nova. Blue Singles™ Competent Cells and ampicillin or chloramphenicol selection. Giga kits, which use Nova. Blue Giga. Singles™ Competent Cells, require one hour for outgrowth.
  • References
  • Novy, R.E., et al. 1996. in. Novations 6 , 7. Clark, J.M. 1988. Nucleic Acids Res . 16 , 9677. Brownstein, J.M., et al. 1996. Bio. Techniques 20 , 1004. Magnuson, V.L., et al. 1996. Bio. Techniques 21 , 700. Hu, G. 1993. DNA and Cell Biology 12 , 763.
  • Components
  • Quality Level MQ100
  • 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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