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Rapid DNA Ligation Kit


The Rapid DNA Ligation Kit enables fast sticky-end or blunt-end DNA ligation in only 5 minutes at room temperature.
  
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The Thermo Scientific Rapid DNA Ligation Kit enables fast sticky-end or blunt-end DNA ligation in only 5 minutes at room temperature (see Figure 1 in Supportig Data).

The kit contains T4 DNA ligase and a specially-formulated 5X rapid ligation buffer optimized for fast and efficient DNA ligation. Fast ligation efficiency is equal to that obtained with T4 DNA ligase in a standard 1 hour ligation. The ligation reaction mixture can be used directly for bacterial transformation with the TransformAid Bacterial Transformation Kit or with other conventional transformation procedures.

Highlights

  • Fast – ligation of either sticky- or blunt-end DNA in only 5 minutes
  • Convenient – reaction mixture can be used directly for bacterial transformation

Applications

  • Routine cloning experiments
  • Blunt-end cloning
  • Library construction
  • TA cloning

Includes

Note

Prior to electroporation, it is necessary to inactivate T4 DNA ligase (see Reference 1) by chloroform extraction or spin column purification, e.g. with Thermo Scientific GeneJET PCR Purification Kit (K0701).

  
Storage Condition-20 C
HazardousNo
Quality Control
  • The kit is tested in ligation of a blunt-end 2.3 kb DNA fragment into a pUC19 DNA/SmaI dephosphorylated cloning vector.
  • Typical yield of white colonies after transformation of competent E. coli XL1-Blue cells is more than 1 x 105 per µg DNA with typical efficiency of competent cells of 107 transformants per µg of supercoiled DNA.
Ligation of sticky-, blunt-end DNA fragments with the Rapid DNA Ligation Kit

Ligation of sticky-, blunt-end DNA fragments with the Rapid DNA Ligation Kit

Ligation of sticky-, blunt-end DNA fragments with the Rapid DNA Ligation Kit

Ligation of sticky- and blunt-end DNA fragments with the Rapid DNA Ligation Kit. Blunt-end ligation. pUC19 DNA/SmaI dephosphorylated vector was ligated with Lambda DNA/PvuII fragment (2.3 kb). Sticky-end ligation. pUC19 DNA/PstI dephosphorylated vector was ligated with Lambda DNA/PstI fragment (2.1 kb). The efficiency of ligation was determined by transformation of E. coli XL1-Blue cells.


References

  1. B. K. Michelsen, Transformation of Escherichia coli increases 260-fold upon inactivation of T4 DNA ligase. Anal. Biochem. 225, 172 (1995).

Protocols and Product Manuals

pdf   Rapid DNA Ligation Kit - Product Information