DNA Ligation Kit

• DNA Ligation Kit is a two-component system that allows rapid performance of all cloning procedures requiring the covalent joining of DNA molecules. The kit consists of a T4 DNA Ligase enzyme mix and a reaction buffer T4 DNA Ligase catalyzes the bonding of sticky or blunt-ended double-stranded DNA or RNA between the 5'-P end and the 3'-OH end with a phosphodiester bond. At the same time, the enzyme can also repair single-stranded nicks in double-stranded DNA, RNA, and DNA/RNA hybrid chains.

• Activity definition: One unit of enzyme can catalyze 1nmol of 32P-labeled pyrophosphate into ATP through displacement reaction within 20 minutes at 37°C (one unit is equal to about 200 sticky-end attachment units).

• Kit components:

1. T4 DNA Ligase Mix [5U/ul] (250U / 500U / 1000U)
2. T4 DNA Ligase Buffer [5x] (1ml / 2 x 1ml / 4 x 1ml)

• Buffer composition:

Storage buffer: 20mM Tris-HCl (pH 7.5), 50mM KCl, 1 mM DTT, 50% (v/v) glycerol
Ligase Buffer [5x]: 250mM Tris-HCl (pH 7.6), 50mM MgCl2, 5mM ATP, 5mM DTT, Enhancer