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TypeOne Restriction Inhibitor

TypeOne Restriction Inhibitor

Increase plasmid transformation efficiency in bacteria, and insertion efficiency of EZ-Tn5 Transposome complexes.
  • Better transformation efficiencies with restriction system inhibition
  • Increase the insertion efficiency of EZ-Tn5 Transposome complexes.
  • Enhance the transformation efficiency of unmodified DNA by blocking type I R-M systems, which are widespread in Eubacteria and Archaebacteria.

Size: 100 µg

Item ID TY0261H
TBD
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Product information

DNA transformation can be difficult to achieve in many bacterial strains due to the presence of one or more restriction and modification (R-M) systems that cleave unmodified DNA that is recognised as "foreign." TypeOne™ Restriction Inhibitor* provides a powerful method for increasing transformation efficiencies in bacterial strains with type I R-M systems. Developed as a unique preparation of a phage protein called ocr, 1 TypeOne Inhibitor is readily electroporated into cells along with transforming DNA. in vivo, the protein acts as a molecular decoy that blocks the DNA-binding site of type I R-M enzymes and inhibits DNA cleavage.

Applications

  • Increase the transformation efficiency of unmodified DNA 2 by blocking type I R-M systems, which are widespread in Eubacteria and Archaebacteria. 3
  • Increase the insertion efficiency of EZ-Tn5™ Transposome™ complexes. 2

Benefits

  • Easy to use: Simply add TypeOne Restriction Inhibitor to unmodified DNA or a Transposome and electroporate using standard methods.
  • Blocks all known families of type I R-M enzymes 1 and can therefore be used to increase transformation efficiencies in a variety of bacterial cells.

Table 1. Effect of TypeOne™ Restriction Inhibitor on transformation efficiencies.

Strain (Type I R-M system)

TypeOne™
Inhibitor
Type of DNA or Transposome™ Recombinants
per µg DNA
S. typhimurium LT2 ( StyL TIII) pUC19 (100 pg) 3.0 × 10 6
S. typhimurium LT2 ( StyL TIII) X pUC19 (100 pg) 3.0 × 10 8
E. coli MG1655 ( EcoK1) 48 kb fosmid (50 ng) 3.0 × 10 3
E. coli MG1655 ( EcoK1) X 48 kb fosmid (50 ng) 1.4 × 10 6
S. typhimurium LT2 ( StyL TIII) EZ-Tn5™ <R6Kγ ori/KAN-2>Tnp Transposome™ (1 µl) 1.3 × 10 4
S. typhimurium LT2 ( StyL TIII) X EZ-Tn5™ <R6Kγ ori/KAN-2>Tnp Transposome™ (1 µl) 1.0 × 10 6

References

  1. Walkinshaw, M.D. et al. (2002) Molec. Cell 9, 187.
  2. Hoff man, L. et al. (2002) Epicentre Forum 9(2), 8.
  3. Murray, N.E. et al. (2000) Microbial. Molec. Biol. Rev. 64, 412.

*Covered by issued and/or pending patents, exclusively licensed or assigned to Epicentre® (an Illumina® Company).

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