T4PNK-RO

T4 Polynucleotide Kinase, 3′-phosphatase free

from phage T4 am N81 pse T1 infected Escherichia coli BB

• 別名: kinase, t4 polynucleotide, 3′-phosphatase-free
• UNSPSCコード: 12352200

特性

• フォーム: solution
• 比活性: ≥40 x 10³ units/mg protein; 40000 U/mg
• 包装: pkg of 1,000 U (10838292001); pkg of 200 U (10709557001)
• メーカー／製品名: Roche
• 保管温度: −20°C

詳細

詳細

T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate group of ATP to the 5′-hydroxyl terminus of DNA or RNA. It also can catalyze the exchange of 5′-terminal phosphate groups. In contrast to the wild type E. coli enzyme, this phage derivative lacks 3′-phosphatase activity. The assay time is 30 minutes and volume activity is 10000 U/ml.

アプリケーション

Use T4 Polynucleotide Kinase, 3′-phosphatase-free, to:

• Phosphorylate the 5′ ends of RNA or DNA
• Phosphorylate the 3′ ends of RNA to prevent cyclization
• Add a 5′[32P]-terminal label to 3′-CMP, forming 5′[32P]pCp. This substrate is commonly used for 3′-end labeling of RNA with T4 RNA ligase.
• Label 5′-hydroxyl ends of DNA with [?-32P]-ATP (either by direct phosphorylation or by phosphate exchange)
• Phosphorylate the 5′-hydroxyl ends of oligonucleotides obtained from automated synthesizers
• Phosphorylate the 5′-hydroxyl ends of linkers.
• Purify recombinant RNA in E. coli.

Note: The 3′-phosphatase activity of the wild-type kinase is not observed under optimal incubation conditions with the mutant T4 Polynucleotide Kinase.

生物化学的／生理学的作用

T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate group of ATP to the 5′-hydroxyl terminus of DNA or RNA. It also can catalyze the exchange of 5′-terminal phosphate groups. In contrast to the wild type E. coli enzyme, this phage derivative lacks 3′-phosphatase activity. The assay time is 30 minutes and volume activity is 10000 U/ml.

包装

1 kit containing 2 components

品質

Absence of 3′-phosphatase, endonucleases, nicking activity, 3′-exonucleases, and ribonucleases, tested according to the current Quality Control procedures; function tested using the DNA 5′-End Labeling Kit.

単位の定義

One unit is the enzyme activity which catalyzes the incorporation of 1 nmol ³²P into acid-precipitable products within 30 minutes at +37 °C.

Volume Activity: 10 x 10³ U/ml

その他情報

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

キットの構成要素のみ

• Enzyme Solution, in buffer, pH 7.5 (+25 °C) 10 U/μl
• Phosphorylation Buffer 10x concentrated

安全性情報

• 保管分類コード: 12 - Non Combustible Liquids
• WGK: WGK 1
• 引火点(℃): does not flash