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- http://lucigen.com/store/docs/manuals/MA069-T4-polynucleotide-kinase.pdf
- 30061-1
- 2026年01月08日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20º C
- 英文名：
T4 Polynucleotide Kinase
- 库存：
大量
- 供应商：
中北林格
- 规格：
1,000 U
T4 Polynucleotide Kinase
- Recombinant T4 Polynucleotide Kinase (T4 PNK).
- Applications include end-labeling and insert preparation for cloning.
T4 Polynucleotide Kinase catalyzes the transfer and exchange of gamma-phosphate of ATP to the 5´-hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates.
Source:
Purified from a strain of E. coli that carries a T4 Polynucleotide Kinase expression plasmid.
Applications:
- End-labeling DNA for probes and DNA sequencing (1)
- Addition of 5´-phosphates to oligonucleotides to allow subsequent ligation (2)
Heat inactivation: 70°C for 15 minutes.
Storage:
Enzyme and buffer should be stored at -20°C. Enzyme is stable for 12 months if handled properly.
Additional Information:
- Ammonium ions are potent inhibitors of T4 Polynucleotide Kinase. DNA should not be precipitated from or dissolved in buffers containing ammonium salts.
- Protruding 5’-overhangs are more efficiently phosphorylated than blunt- or recessed 5’ overhangs.
References:
- Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 9.55-9.56, 9.68-9.69.
- Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 13.16-13.17.
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文献和实验Polynucleotide Kinase (EC 2.7.1.78)
The enzyme polynucleotide kinase (PNK, or ATP:5′-dephospho-polynucleotide 5′-phosphatase) catalyzes the transfer of a γ-phosphate group from a 5′-nucleoside triphosphate moiety to a free 5′-hydroxyl of a polynucleotide such as DNA or RNA, to form
Radiolabeling of DNA Using Polynucleotide Kinase
Labeling of DNA fragments at the 5′ end with polynucleotide kinase is one of the methods currently employed to obtain highly labeled DNA for sequencing purposes. This end-labeled DNA is used in the partial chemical degradation method
由 T偶数噬菌体所感染的大肠杆菌分离出来的酶，能催化 ATP的γ -磷酸转移到 DNA或 RNA的 5′ -OH末端生成 ADP的反应。 EC2． 7． 1． 78。因为此酶的催化反应特异性很高，可用 32 P标记的 ATP特异地标记多核苷酸的 5′末端。广泛应用于多核苷酸的链长的测定和 5′末端核苷酸排列的确定等核酸结构的研究。
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