描述

Thermo Scientific Rapid DNA Ligation Kit enables fast sticky-end or blunt-end DNA ligation in only 5 minutes at room temperature.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

• T4 DNA Ligase
• 5X Rapid Ligation Buffer
• Water, nuclease-free
• Detailed Protocol

Note

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

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Rapid DNA Ligation Kit

Ligation of sticky-, 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.