NEB M0202S T4 DNA Ligase 20000U

feature

Efficient connection between adhesive end and flat end
T/A cloning
DsDNA cleavage repair
Constructing a high abundance clone library
The connection between RNA and DNA

NEB M0202S T4 DNA Ligase 20000U

Overview

This enzyme catalyzes the formation of phosphodiester bonds between adjacent 5 '- phosphate and 3' - hydroxyl termini on double stranded DNA or RNA. This enzyme can not only catalyze the connection between flat or sticky ends, but also repair single strand breaks in double stranded DNA, RNA, or DNA/RNA hybrid double strands.

source

Purification fromE. coliC600 pcl857 pPLc28 lig8。

reaction conditions

1X T4 DNA Ligase Reaction Buffer
[50 mM Tris HCl (pH 7.5 @ 25 ℃), 10 mM MgCl2, 10 mM DTT, 1 mM ATP]. *DNA 5 'end concentration is 0.1-1.0 μ M, incubated at 16 ℃.
Thermal deactivation: 65 ℃ for 10 minutes.

Unit definition (viscous terminal active unit)

1 unit refers to the amount of enzyme required to ligate 50% of HindIII digested lambda DNA fragments [5 'end concentration of 0.12 μ M (300 μ g/ml)] in 20 μ l 1X T4 DNA ligase reaction buffer at 16 ℃ for 30 minutes.

concentration

400000 units/ml and 2000000 units/ml.

Precautions

WithE. coliDNA ligases require NAD+as a cofactor, while T4 DNA ligases require ATP as a cofactor.

The diluted T4 DNA ligase should be stored at -20 ℃ in 50% glycerol storage buffer (dilution buffer A, NEB # B8001). If used immediately after dilution, it can also be diluted with 1X T4 DNA ligase reaction buffer.

As long as 1 mM ATP is added to the reaction system, the ligation reaction can be carried out in the four restriction enzyme buffers provided by NEB or in

T4 DNA Polynucleotide Kinase Reaction Buffer.

T4 DNA ligase

Item Number

specification

Price

Beijing inventory

Shanghai inventory

#M0202L

100,000 units

have

have

#M0202M

(High concentration 5X) 100000 units

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have

#M0202S

20,000 units

have

have

#M0202T

(High concentration 5X) 20000 units

have

have

#M0202V

10,000 units

have

have