T4 Polynucleotide Kinase
(Synonyms: T4多聚核苷酸激酶) 目录号 : GC26293T4 Polynucleotide Kinase,简称T4 PNK,中文名称T4多聚核苷酸激酶,是一种多聚核苷酸5'羟基激酶,
Sample solution is provided at 25 µL, 10mM.
T4 Polynucleotide Kinase, abbreviated as T4 PNK, is a polynucleotide 5 'hydroxy kinase that catalyzes ATP γ The phosphate group is transferred to the 5 'hydroxyl group of single or double stranded DNA, RNA, oligonucleotides, or single nucleotides with a 3' phosphate group. Other NTPs can also produce the same reaction: 5 '- OH+NTP → 5' - P+NDP.
T4 Polynucleotide Kinase,简称T4 PNK,中文名称T4多聚核苷酸激酶,是一种多聚核苷酸5'羟基激酶,可以催化ATP的γ位磷酸基团向单链或双链DNA、RNA、寡核苷酸或带有3'磷酸基团的单核苷酸的5'羟基转移。其他NTP也可产生相同的反应:5'-OH + NTP → 5'-P + NDP。
操作流程
反应体系的设置:
1) 溶解并混匀PCR反应所需的各种溶液。并放置于冰浴上或冰盒内。建议反应PCR液体分装使用,避免反复冻融。
2) 参考下表设置的反应,建议PCR反应体系的配置在冰浴或在冰盒上进行:
试剂 | 50 μl 反应体系 |
T4 Polynucleotide Kinase (10 U/ul) | 1-2 μl |
5×T4 PNK Buffer | 10 μl |
待磷酸化DNA | — |
γ-[32P]-ATP/ATP(10mM) | 5 μl |
T4PNK、模板、ATP可根据实验具体调整用量,不足50 μl 用RNaseFree-H2O 补足 | |
体积 | 50 μl |
3) 37°C温育30分钟。
Components | GC26293-0500 | GC26293-1000 |
T4 Polynucleotide Kinase(10 U/ul) | 50 µl | 100 µl |
5X T4 PNK Buffer | 500 µl | 1 ml |