Merck
所有图片(1)

D8276

Sigma-Aldrich

DNA 聚合酶 I, Klenow 片段 来源于大肠杆菌

buffered aqueous glycerol solution

CAS号:
MDL编号:
NACRES:
NA.53

等级

for molecular biology

形式

buffered aqueous glycerol solution

分子量

103 kDa

浓度

~3,000 units/mL

UniProt登记号

异质活性

Endonuclease, none detected

运输

wet ice

储存温度

−20°C

Gene Information

Escherichia coli K12 ... polA(948356)

一般描述

DNA 聚合酶I 经蛋白酶消化产生两个断片(小断片和大断片)。大断片(Klenow 断片)丢失存在于完整全酶中的5′核酸外切酶活性。然而,它保留了聚合酶 5′→3 ′活性和天然酶的 3′→5 ′核酸外切酶活性。

应用

适用于:
  • DNA 测序(Sanger 双脱氧法)
  • cDNA 互补链的合成
  • 将5′ -突出部分填入双链 DNA 形成钝端
  • 采用寡核苷酸通过第二链合成进行DNA诱变
  • 采用随机引物法标记 DNA

组分

DNA 聚合酶 I 以溶于 50 mM Tris-HCl (pH 7.5)、1 mM EDTA、5 mM 二硫苏糖醇和 50% 甘油 (v/v) 的溶液形式提供。

单位定义

在 37 °C下,一个单位在30分钟内将 10 毫微摩尔的脱氧核苷三磷酸转化为酸不溶性物质。

重悬

酶溶液可用 50 mM Tris-HCl (pH 7.5)、100 mM 硫酸铵、10 mM 2-巯基乙醇和 1 mg/ml牛血清白蛋白稀释。

分析说明

在含 50 mM 磷酸钾 (pH 7.5)、3 mM MgCl 2 、1 mM 2-巯基乙醇、32.5 μM 32P-dATP、32.5μM dTTP、62.5μg/ml poly (dA-dT) 和 0.01-1单位酶。

储存分类代码

12 - Non Combustible Liquids

WGK

WGK 2

闪点(F)

Not applicable

闪点(C)

Not applicable

个人防护装备

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

分析证书

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原产地证书 (CofO)

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Quotes and Ordering

Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
Methods in enzymology, 152, 330-335 (1987-01-01)
H Klenow et al.
Proceedings of the National Academy of Sciences of the United States of America, 65(1), 168-175 (1970-01-01)
Purification of DNA polymerase from E. coli B has in two cases each time led to the isolation of two separate polymerase activities, enzyme A and enzyme B. Enzyme A was in contrast to enzyme B almost completely devoid of
L M Houdebine
Nucleic acids research, 3(3), 615-630 (1976-03-01)
E.Coli DNA polymerase I (Klenow subfragment) was used for the synthesis of complementary DNA with the mRNAs for rabbit milk proteins as templates. The cDNA formed, contained 200 nucleotides and represented about 20% of the mRNA template. The cDNA was
R B Wallace et al.
Science (New York, N.Y.), 209(4463), 1396-1400 (1980-09-19)
Many eukaryotic genes contain intevening sequences, segments of DNA that interrupt the continuity of the gene. They are removed from RNA transcripts of the gene by a process known as splicing. The intervening sequence in a yeast tyrosine transfer RNA
DNA polymerase versus DNA binding to the anticancer drug, cis-platin.
Bose, R.N., et al.
Inorgorganica Chimica Acta, 300, 937-937 (2000)

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