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Merck
CN

P4390

Sigma-Aldrich

Polynucleotide Kinase from T4-infected Escherichia coli

10 units/μL, buffered aqueous glycerol solution

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About This Item

CAS Number:
MDL编号:
UNSPSC代码:
12352204
NACRES:
NA.53

等级

Molecular Biology

质量水平

表单

buffered aqueous glycerol solution

分子量

33 kDa

浓度

10 units/μL

异质活性

Endonuclease and exonuclease, none detected

运输

wet ice

储存温度

−20°C

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应用

Suitable for:
  • Sequencing or nucleic acid tagging (DNA and RNA) by 5′-end labeling
  • 5′ phosphorylation of oligonucleotides
  • Removal of 3′-phosphate groups from phosphorylpolynucleotides

组分

T4 Polynucleotide Kinase is supplied in a solution of 50% glycerol (v/v), 20 mM Tris-HCl (pH 7.5), 25 mM KCl, 2mM DTT, 0.1 mM EDTA, and 0.1 μM ATP.

原理

Polynucleotide kinase catalyses a "forward reaction" transfer of the γ-phosphate of ATP to the 5′ hydroxyl terminus of single- and double-stranded nucleic acids (DNA and RNA) and 3′-nucleoside monophosphates. In exchange reactions containing ADP, the enzyme will catalyze the exchange of 5′-terminal phosphate groups and ATP. The 3′-phosphatase activity enables the enzyme to remove 3′-phosphoryl groups from phosphorylpolynucleotides.
1. Forward reaction: Transfer of the labeled γ-phosphate from [γ-32P]-ATP to the free 5′-hydroxyl group of the substrate.
5′-HO-DNA + [γ-32P]-ATP → 5′-32PO-DNA + ADP.
Substrates that do not have a free 5′-hydroxyl require prior dephosphorylation by alkaline phosphatase.
2. Exchange reaction: First, the terminal 5′-phosphate is transferred from the substrate to ADP present in the reaction mixture. Then, the labeled γ-phosphate from [γ-32P]-ATP is transferred to the free hydroxyl group of the substrate.
5′-PO-DNA + ADP → 5′-HO-DNA + ATP
5′-HO-DNA + [γ-32P]-ATP → 5′-32PO-DNA + ADP

单位定义

One unit catalyzes the transfer of one nanomole of 32P to the 5′-end of micrococcal nuclease-treated DNA in 30 min. at 37 °C. Transfer is detected as incorporation into acid-insoluble material.

分析说明

Activity is determined in a reaction mixture containing 40 mM Tris-HCl (pH 7.5), with 10 mM MgCl2, 5 mM dithiothreitol, 0.5 mM 5′-OH polynucleotide ends, and mM [γ-32P]-ATP.

象形图

Health hazard

警示用语:

Danger

危险声明

预防措施声明

危险分类

Resp. Sens. 1

储存分类代码

10 - Combustible liquids

WGK

WGK 3

闪点(°F)

Not applicable

闪点(°C)

Not applicable

个人防护装备

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


历史批次信息供参考:

分析证书(COA)

Lot/Batch Number

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