Manual

Product Number: LG01

Shipping and Storage

Stored at -20ºC.

Components

Component	LG01 40KU
T4 DNA Ligase (1000U/µl)	40KU
10×T4 Reaction Buffer	0.3ml

Description

T4 DNA ligase can catalyze the phosphodiester bond between the 5 '- P and 3' - OH ends of sticky or flat double stranded DNA or RNA. This catalytic reaction requires ATP as a cofactor. At the same time, T4 DNA ligase can repair single strand defects on double stranded DNA, double stranded RNA, or DNA/RNA hybrids.

Purity: Free of DNA endonucleases, exonucleases, and phosphatases, free of RNAses, meeting the requirements of conventional ligation reactions.

Storage solution:20 mM Tris, pH 7.5, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA and 50% (v/v) glycerol。

10X Ligation Buffer: 400 mM Tris, pH 7.8, 100 mM MgCl₂,100 mM DTT, 10 mM ATP。

Inactivation or inhibition: Incubation at 65ºC for 10 minutes can lead to inactivation of T4 DNA Ligase; When the concentration of NaCl or KCl is greater than 200mM, it strongly inhibits T4 DNA Ligase.

The effect of coating LB plates with T4 DNA Ligase linked products after transformation into competent states is shown in Figure 1.

The LB plate effect obtained by coating the T4 DNA Ligase ligation product onto competent bacteria. A. The carrier after double enzyme digestion was self linked overnight; B. The vector after double enzyme digestion is connected overnight with the fragment to be inserted.

Source

This T4 DNA ligase is expressed by Escherichia coli, and the source of the expressed gene is T4 thermophilic bacteria.

Application

T4 DNA ligase is commonly used for connecting DNA fragments to vectors, linkers, or adapters. It can also be used for defect repair and Ligase mediated RNA detection.

Unit definition

One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min at 16ºC in 20µl of the assay mixture containing 50mM Tris, pH 7.5, 10mM MgCl₂, 10mM DTT, 1 mM ATP, 25µg/ml BSA and a 5'-DNA termini concentration of 0.12µM (300µg/ml)。200U is equal to 1 Weiss unit, and based on Weiss unit, this product has a total of 200 units.