QuickLink DNA Ligation Kit, joins blunt-end and sticky-end DNA fragments

Application

QuickLink^™ DNA Ligation Kit has been used for the ligation of pGL2-basic luciferase-reporter plasmid with vascular endothelial growth factor (VEGF) promoter sequence and for the ligation of NLR family caspase activating and recruitment domain (CARD)-containing protein 4 (Nlrc4) promoter with the luciferase plasmid vector pGL4.10-luc2

The QuickLink^™ DNA Ligation Kit is suitable for: Joining blunt or cohesive-end fragments of DNA into a cloning vector Recircularization of linear DNA Formation of concatamers dsDNA nick repair

Components

Sufficient for 50 reactions: 500uL 2X Ligation Buffer A (L9537) 100uL 5X Ligation Buffer B (L9662) 250 units T4 DNA Ligase (D2886) in 50% glycerol with 10 mM Tris-HCl (pH 7.5) 50 mM KCl, and 1 mM DTT

Features and Benefits

Fast 5 minutes ligation High ligation efficiency Room temperature reactions – no cooling required Perform bacterial transformation with the reaction mixture

General description

Sigma’;s DNA Ligation Kit contains all of the reagents necessary to perform DNA ligation reactions at room temperature using blunt or sticky ends. This kit replaces previous workflows requiring cooking steps and long incubations. Pre-made buffers allow for fast & easy setup.

Legal Information

QuickLink is a trademark of Sigma-Aldrich Co. LLC

Principle

One of the most important steps in the cloning process is the ligation of linear DNA into a cloning vector. DNA ligations are performed by incubating DNA fragments with appropriately linearized cloning vectors in the presence of buffer, ATP, and DNA ligase. Many parameters affect ligations such as the relative ratio of insert to vector, the quality and type of the DNA ends, the temperature of ligation and the concentration of DNA.