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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20°C
- 库存:
100
- 供应商:
ABclonal
- 规格:
200 U/1000 U/5000 U
| 规格: | 200 U | 产品价格: | ¥299.0 |
|---|---|---|---|
| 规格: | 1000 U | 产品价格: | ¥719.0 |
| 规格: | 5000 U | 产品价格: | ¥2169.0 |
产品描述
Klenow片段(3'-5' exo-)是N-末端截短型的DNA聚合酶I,保留DNA聚合酶活性,缺失5'-3'外切酶活性,同时因D355A和E357A突变而缺失3'-5'外切酶活性。Klenow片段(3'-5' -)聚合酶适用于随机引物标记、Sanger法DNA测序、cDNA的二链合成和突变实验的二链合成。
产品组分
| Klenow Fragment 3’→5’ exo- (5,000 U/ml) | RM20516 |
| 10X ABuffer B | RM20126 |
产品信息
| 产品来源 | 携带截短323个密码子的E.coli polA基因(D355A, E357A)在大肠杆菌中诱导重组表达和分离纯化得到。 |
|---|---|
| 应用 | A-tailing, DNA聚合酶扩增反应, DNA测序,RT-PCR和cDNA合成,PCR |
| 活性定义 | 1活性单位(U)指在37°C 30 min内,将10 nmol脱氧核苷酸掺入到酸不溶性物质所需的酶量。 |
| 保存条件 | 25 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4 @25℃ |
| 保存温度 | -20°C |
| 反应条件 | 1x ABuffer B, 37°C孵育 |
| 反应缓冲液或预混液 | 1X ABuffer B: 10 mM Tris-HC, 50 mM NaCl, 10 mM MgCl2, 1 mM DTT, pH7.9 @ 25°C |
| 热失活 | 75°C 加热 20 min |
| 5' - 3' 核酸外切酶 | 无 |
| 3' - 5' 核酸外切酶 | 无 |
| 链置换活性 | +++ |
| 蛋白分子量 | 68 kDa |
| 出错率 | ~ 100 x 10-6 bases |
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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
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