DNA Polymerase I Large (Klenow) Fragment

产品编号：M2201 品牌：promega 原厂货号：M2201
产品分类：克隆 > 聚合酶 > DNA聚合酶
应用分类：克隆及合成生物学 > 平端克隆

包装：

150u

运保温度:

-20℃

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其它组分：

DNA Polymerase 10X Buffer【子货号：M195A，包装：1 x 1ml，，运保温度：-20℃】

DNA Polymerase I Large (Klenow) Fragment【子货号：M220A，包装：1 x 150u，，运保温度：-20℃】

描述：

说明

DNA Polymerase I Large (Klenow) Fragment (DNA 聚合酶I 大(Klenow) 片段) 是DNA 依赖的DNA 聚合酶，它没有完整的E. coli DNA Polymerase I 所具有的 5` → 3` 外切酶活性，但保留了5` → 3` 聚合酶、3` → 5` 外切酶及链置换活性。该酶是DNA 聚合酶I 的C- 末端片段，68kDa。Klenow 片段5` → 3` 聚合酶活性经常被用于以非标记的或标记的dNTPs 补平5`- 突出端、单链或双链DNA 模板测序、合成寡核苷酸进行体外突变、cDNA第二链合成以及制备单链DNA探针。3` → 5` 外切酶活性可用于从3`- 突出端产生平末端。

特点

• 灵活性：DNA Polymerase I Large (Klenow) Fragment 可用于多种分子实验。在多种Promega 1× 内切酶缓冲液中它都能保持活性。

• 高温灭活：DNA Polymerase I Large (Klenow) Fragment 在75℃加热10 分钟失活。

• 提供10× 反应缓冲液：500mM Tris-HCl (pH 7.2，25℃ )，100mM MgSO₄，1mM DTT。

• 可定制或批量订购：从以下网址获得更多关于该产品的定制选项：www.promega.com/myway/。

原厂资料：

DNA Polymerase I Large (Klenow) Fragment is a DNA-dependent DNA polymerase that lacks the 5´→3´ exonuclease activity of intact E. coli DNA Polymerase I but retains its 5´→3´ polymerase, 3´→5´ exonuclease and strand displacement activities. The enzyme is a 68kDa C-terminal fragment of DNA Polymerase I. The 5´→3´ polymerase activity of Klenow Fragment can be used to fill in 5´-protruding ends with unlabeled or labeled dNTPs, to sequence single- or double-stranded DNA templates, for in vitro mutagenesis using synthetic oligonucleotides, for cDNA second-strand synthesis and to generate single-stranded DNA probes. The 3´→5´ exonuclease activity can be used to generate blunt ends from a 3´-overhang.

Features - Benefits

Flexible: DNA Polymerase I Large (Klenow) Fragment may be used in a variety of molecular applications. It is also active in many Promega 1X restriction enzyme buffers.
May Be Heat-Inactivated: DNA Polymerase I Large (Klenow) Fragment is inactivated by heating at 75°C for 10 minutes.
Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C), 100mM MgSO4, 1mM DTT.
Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

Applications

Filling-in or labeling 3´ ends of 5´ overhanging double-stranded DNA.
Second-strand cDNA synthesis.
DNA sequencing by the dideoxy method.

References

Anderson, S. et al. (1980) Nucl. Acids Res. 8, 1731–43.
Sanger, F. et al. (1977) Proc. Natl. Acad. Sci. USA 74, 5463–7.
Wallace, R.B. et al. (1980) Science 209, 1396–400.
Houdebine, L.M. (1976) Nucl. Acids Res. 3, 615–30.
Feinberg, A.P. and Vogelstein, B. (1983) Anal. Biochem. 132, 6–13.
Tabor, S. and Struhl, K. (1987) In: Current Protocols in Molecular Biology, Ausubel, F.M. et al., eds., John Wiley and Sons, New York, NY.
Joyce, C.M. and Grindley, N.D.F. (1983) Proc. Natl. Acad. Sci. USA 80, 1830–4.