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- 文献和实验
- 技术资料
- 英文名：
T4 Polynucleotide Kinase (10 U/µL)
- 规格：
2,500 units
描述
Thermo Scientific™ T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of the gamma-phosphate from ATP to the 5'-OH group of single- and double-stranded DNAs and RNAs, oligonucleotides, or nucleoside 3'-monophosphates (forward reaction). The reaction is reversible. In the presence of ADP, T4 Polynucleotide Kinase exhibits 5'-phosphatase activity and catalyzes the exchange of phosphate groups between 5'-P-oligo-polynucleotides and ATP (exchange reaction). The enzyme is also a 3'-phosphatase.Highlights
• Active in Thermo Scientific restriction enzyme, RT, and T4 DNA ligase buffers
Applications
• Labeling 5' -termini of nucleic acids to be used as:
--Probes for hybridization
--Probes for transcript mapping
--Markers for gel electrophoresis
--Primers for DNA sequencing
--Primers for PCR
• 5'-phosphorylation of oligonucleotides, PCR products, other DNA or RNA prior to ligation
• Phosphorylation of PCR primers
• Detection of DNA modification by the [32P]-postlabeling assay
• Removal of 3'-phosphate groups
Notes
• The 5'-termini of nucleic acids can be labeled by either the forward or the exchange reaction.
• Polyethylene glycol (PEG) and spermidine improve the rate and efficiency of the phosphorylation reaction. PEG is used in the exchange reaction mixture.
• Since T4 Polynucleotide Kinase is inhibited by ammonium ions, use sodium acetate to precipitate DNA prior to phosphorylation.
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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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