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DNALIG-RO

Roche

T4 DNA Ligase

solution, from Escherichia coli(NM 989), suitable for activity assay

Synonym(s):
dna ligase, t4

biological source

Escherichia coli (NM 989)

Quality Level

recombinant

expressed in E. coli

description

recombinant form of the enzyme from T4 phage

form

solution

specific activity

~3000 units/mg protein

packaging

pkg of 100 U (10481220001 [1 U/μl])
pkg of 500 U (10716359001 [1 U/μl])
pkg of 500 U (10799009001 [5 U/μl])

manufacturer/tradename

Roche

parameter

4-22 °C optimum reaction temp.

technique(s)

activity assay: suitable

optimum pH

7.2-7.8

storage temp.

−20°C

General description

T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighboring 3′-hydroxyl- and 5′-phosphate ends in double-stranded DNA. Single-stranded nicks in double-stranded DNA are also closed by T4 DNA Ligase.

Contents
T4 DNA Ligase, supplied with 10x concentrated ligation buffer that includes ATP.

Specificity

Heat inactivation: The enzyme can be inactivated by heating (65 °C, 10 minutes).

Application

Ligation of sticky- and blunt-ended DNA fragments.

Quality

Quality

Tested for the absence of deoxyribonucleases and exonucleases, according to the current Quality Control procedures.

Unit Definition

One unit of T4 DNA ligase is the amount of enzyme activity that converts 1 nmol 32P from pyrophosphate into Norit-absorbable material in 20 minutes at +37 °C. One unit corresponds to 0.2 U determined in the exonuclease III resistance assay. Dilution buffer: 50 mM Tris-HCl, 10 mM dithioerythritol, bovine serum albumin, 500 μg/ml, pH 7.6 (+25 °C).

Volume Activity: 1 x 103 U/ml; 5 x 103 U/ml

Preparation Note

Activator: Mg2+ (10mM), DTT
Working solution: For dilution of the enzyme Roche recommends using a buffer containing the components of the storage buffer: 20 mM Tris-HCl, 60 mM KCl, 1 mM EDTA, 5 mM dithoerythritol, 50% glycerol (v/v), pH 7.5 (4 °C).

Other Notes

For life science research only. Not for use in diagnostic procedures.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

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Certificate of Origin

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