Quick Ligase (EN12)

Quick Ligase (EN12)

125,00 302,00 

ATP-dependent recombinant T4 DNA ligase for efficient ligation of cohesive and or blunt end DNA fragments in 5 and 15 minutes respectively.
EN12, EN12-050, EN12-150

SKU: EN12 Category:

Available Options:

Reactions

50 reactions, 250 reactions

Product Information

Quick Ligase is an ATP-dependent recombinant enzyme isolated from Escherichia coli strain and used to clone DNA in just 5 to 15 minutes. Quick Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA, as well as repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

Applications:

  • Ligation in just 5 – 15 minutes
  • Cloning PCR products
  • Cloning restriction fragments
  • Joining double-stranded oligonucleotide linkers or adaptors to DNA
  • Site-directed mutagenesis
  • Nick repair in duplex DNA, RNA or DNA/RNA hybrids
  • Self-circularization of linear DNA
  • LM PCR methods (Ligation Mediated PCR), e.g. amplified fragment length polymorphism (AFLP)

Unit definition

One (Weiss) unit of Quick Ligase catalyzes the conversion of 1 nmol of 32P from pyrophosphate into Norit-adsorbable material in 20 minutes at 37°C.

Protocol & Manual

Quick Ligase (EN12) - Manual

Technical Tips

  • The 10x Ligation Buffer and ATP Solution should be thawed and resuspended at room temperature.
  • For blunt-end ligations, use higher quantities of both the vector and the insert DNA.
  • For sticky (cohesive)-end ligations, we recommend heating both the vector and the insert DNA prior to ligation.
  • The electrotransformation efficiency may be improved by heat inactivation of the Quick Ligase and purifi cation of the DNA by means of a spin column purification method (EXTRACTME DNA CLEAN-UP kit or
    EXTRACTME DNA GEL-OUT kit).
  • We recommend using a 3 – 10 molar excess of insert DNA over vector DNA. To calculate the optimal amounts of insert DNA in a ligation reaction,use the following equation:
    Example: If using 20 ng of a vector plasmid (4 kb), for a 5:1 molar ratio of insert : vector, you will require the following quantity of 1 kb insert:
  • The enzyme is inhibited by >200 mM NaCl or KCl concentrations.
  • Inactivate enzyme at 65°C for 10 minutes or at 70°C for 5 minutes.

FAQ

Quality & Safety

Quality control

The product was tested in ligation of HindIII-cut lambda DNA with a different amounts of enzyme. The product is free of unspecific DNA nucleases.

Quick Ligase (EN12) - MSDS


Quick Ligase (EN12) - CoA 1.2018 L.686771

Publications