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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20°C保存。
- 英文名:
DNA Polymerase I Klenow Fragment exo-
- 库存:
充足
- 供应商:
南京诺唯赞生物科技有限公司
- 规格:
5000 U
DNA Polymerase I Klenow Fragment exo-
缺失3’→5’外切酶活性的Klenow片段

双脱氧测序;随机启动标记;建库中3'末端加dA尾

3’末端加dA尾
用随机引物制备探针
随机引物法标记

Klenow片段(3´→5´exo-) 是DNA聚合酶I的N末端截短物,它保留了DNA聚合酶活性,但失去了5´→3´核酸外切酶活性;该酶经突变 (D355A,E357A) 进一步去除了其3´→5´核酸外切酶活性。
贮存缓冲液
25 mM Tris-HCl pH 7.4, 0.1 mM EDTA, 1 mM DTT, 50% glycerol
反应缓冲液 (10 × Blue Buffer)
100 mM Tris-HCl pH 7.9, 500 mM NaCl, 100 mM MgCl2, 10 mM DTT
来源
克隆有来自E.coli DNA Polymerase I Klenow Fragment exo-基因的重组E. coli菌株。
单位定义
37℃、30分钟内使10 nmol的dNTP掺入酸不溶性沉淀物所需要的酶量,定义为1个活性单位 (U)。


反应缓冲液
10 × Blue Buffer
100 mM Tris-HCl pH 7.9 @ 25°C
500 mM NaCl
100 mM MgCl2
10 mM DTT

-20°C保存。
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文献和实验Radiolabeling of DNA with the Klenow Fragment of DNA Polymerase
The Klenow fragment of DNA polymerase I is a proteolytic fragment obtained by the treatment of DNA polymerase I with subtilisin. The fragment still has the polymerase and 3′–5′ exonuclease activity, but lacks the 5′–3′ exonuclease activity
Dideoxy Sequencing Reactions Using Klenow Fragment DNA Polymerase 1
Historically, DNA sequencing by primed synthesis and chain terminators has been performed using the Klenow fragment of DNA polymerase 1(1 . This enzyme was derived from E. coli . DNA Pol I which, in addition to polymerase activity, has both 3′–
Mainly as a result of the booming interest in apoptosis and of the discovery of the occurrence of massive DNA breaks as a major feature of this type of cell death, during the past few years polymerase-based assays for detection of DNA breaks







