重组尿嘧啶DNA糖基化酶 (热敏UDG)

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级别和纯度:

for DNA and RNA applications

1 U/μL

表达系统: E. coli
生物活性: 1 U/μL

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货号 (SKU)	包装规格	是否现货
H292766-100μl	100μL	期货
H292766-100U	100U	期货
H292766-5×100U	5×100U	期货

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PCR (26) RT-qPCR (9) UDG (3) Uracil-DNA Glycosylase (3) 分子生物学用酶 (56)

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基本描述

产品名称

重组尿嘧啶DNA糖基化酶 (热敏UDG)

规格或纯度

无DNA酶和RNA酶, EnzymoPure™, 分子生物学级, for DNA and RNA applications, 重组, 1 U/μL

产品介绍

本公司生产的 Uracil-DNA Glycosylase (Heat-labile, Bacterium)，也称 Heat-labile Bacterial UDG，即热敏型细菌 UDG，来源于嗜冷海洋细菌 (psychrophilic marine bacterium) BMTU3346，可催化含尿嘧啶的 DNA 链中的尿嘧啶 (dU) 碱基和脱氧核糖之间的 N - 糖苷键发生水解，从而释放游离尿嘧啶。Uracil-DNA Glycosylase (UDG) 可以水解含有 dU 的单链或双链 DNA，但不能水解 RNA 或含有 dU 的长度不超过 6 个碱基 DNA 寡聚体。UDG 主要应用于消除 PCR 扩增过程中带来的产物污染问题。其防止污染的原理为：在 PCR 反应中加入适量的 dUTP，以 dUTP 替代 dTTP 掺入 DNA 中，形成含 dU 碱基的 PCR 扩增产物；后续进行 PCR 反应时，使用 UDG 酶选择性切割可能被污染而带入的之前 PCR 扩增产生的含有 dU 的单链或双链 DNA，从而避免之前的 PCR 扩增产物可能的污染对于本次 PCR 扩增带来的负面影响。本产品为热敏型，在 50℃孵育 10 分钟可以迅速不可逆灭活。在 PCR 或 RT-PCR 反应前，PCR 或 RT-PCR 体系中加入 Heat-labile Bacterium UDG 室温处理 10min，即可充分消除可能的之前的含有 dUTP 的 PCR 扩增产物的污染。本产品不仅适用于 PCR，也适用于 qPCR、RT-PCR 和 qRT-PCR 等体系。

来源 (Source)	大肠杆菌重组表达
外观 (Appearance)	无菌液体
保存液 (Storage Buffer)	20mM Tris-HCl (pH 7.5)，50mM NaCl，0.1mM EDTA，1mM DTT，50% (v/v) glycerol
酶浓度 (Enzyme Concentration)	1U/μL
纯度 (Purity)	不含 UDG 酶活力之外的内切或外切脱氧核糖核酸酶、RNase 和磷酸酶活性。
活性定义 (Activity Definition)	One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. Activity is measured by release of [3H]-uracil in a 50 µl Standard Taq Reaction Buffer containing 0.2 µg DNA (104–105 cpm/µg) in 30 minutes at 37℃.

组分和说明

H292766	Component	100U	5×100U	Storage
H292766A	Heat-labile Bacterial UDG (1U/μl)	100µl	5×100µl	-20℃. Avoid freeze/thaw cycle.
H292766B	10× Heat-labile Bacterial UDG Buffer	500µl	5×500µl	-20℃. Avoid freeze/thaw cycle.

产品应用

去除单链或双链 DNA 尿嘧啶碱基；去除含 dU 的 PCR 产物气溶胶污染；在二代测序 (NGS) 应用中，主要用于 mRNA 定向测序文库的构建；用于提高定向诱变方法的效率和用来获得高度标记的寡核苷酸探针；用于 PCR、qPCR、RT-PCR、RT-qPCR 体系。

产品优势

产品为热敏型，在 50℃孵育 10 分钟可以迅速不可逆灭活。

使用说明

1. PCR 反应体系的设置：

a. 按照常规反应体系设置 PCR 或 RT-PCR 反应体系，同时加入 Heat-labile Bacterial UDG 至最终浓度为 0.02U/μl，混匀。通常仅加入 PCR 或 RT-PCR 的 buffer 即可，无需加入 UDG 的 buffer。

b. 25℃孵育 5 - 10min (去除可能的含 dUTP 的 PCR 扩增产物的污染)，随后进入 PCR 或 RT-PCR 程序。

注意事项

(1) Heat-labile Bacterial UDG 酶在大多数 PCR 或 RT-PCR 体系中均具有活性，但对于自行使用的 PCR 或 RT-PCR 体系，首次使用时建议先测试一下是否和所使用的体系兼容。通常取含 dUTP 的 PCR 扩增产物，参考图 1 加入适量 UDG，观察能否有效降解含 dUTP 的 PCR 扩增产物。本产品会被高离子强度 (>100mM) 所抑制。
(2) dNTP/dUTP 推荐选购阿拉丁的D745378 dNTP/dUTP Mixture (2.5mM each/5mM)。
(3) Heat-labile Bacterial UDG 酶对双链 DNA 的活性低于对单链 DNA 的活性，对小的 U-DNA 寡核苷酸和 dUMP 有活性，但对 RNA 或正常的不含 dUTP 的 DNA 没有活性。
(4) Heat-labile Bacterial UDG 酶活对于金属离子没有依赖性，在 Mg²⁺或 EDTA 的存在情况下均具有活性。
(5) 由 Heat-labile Bacterial UDG 酶消化产生的 DNA 链的无碱基位点可通过加热，碱处理或核酸内切核酸酶处理而除去。通常 PCR 反应过程中的加热步骤可以确保 UDG 酶消化的位点被完全剪切开。
(6) Heat-labile Bacterial UDG 酶可以在 PCR 反应前清除不慎污染的含 dUTP 的 PCR 产物，从而避免由于污染导致的 PCR 假阳性结果。
(7) 如果需要将 Heat-labile Bacterial UDG 酶应用于 One-Step RT-PCR 或者 One-Step qRT-PCR 体系，需要选择耐热反转录酶，并需要将逆转录温度设置为 50 - 55℃。
(8) 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
(9) 为了您的安全和健康，请穿实验服并戴一次性手套操作。

The Uracil-DNA Glycosylase (Heat-labile, Bacterium) produced by our company, also known as Heat-labile Bacterial UDG (i.e., heat-labile bacterial UDG), is derived from the psychrophilic marine bacterium BMTU3346. It can catalyze the hydrolysis of the N-glycosidic bond between the uracil (dU) base and deoxyribose in the DNA strand containing uracil, thereby releasing free uracil. Uracil-DNA Glycosylase (UDG) can hydrolyze single-stranded or double-stranded DNA containing dU, but cannot hydrolyze RNA or DNA oligomers containing dU with a length of no more than 6 bases. UDG is mainly used to eliminate product contamination caused during PCR amplification. The principle of preventing contamination is as follows: an appropriate amount of dUTP is added to the PCR reaction, and dUTP is used to replace dTTP for incorporation into DNA to form PCR amplification products containing dU bases; during subsequent PCR reactions, UDG enzyme is used to selectively cleave the single-stranded or double-stranded DNA containing dU from previous PCR amplifications that may be introduced by contamination, thereby avoiding the negative impact of potential contamination from previous PCR amplification products on the current PCR amplification. This product is heat-labile and can be rapidly and irreversibly inactivated by incubation at 50℃ for 10 minutes. Before PCR or RT-PCR reactions, adding Heat-labile Bacterial UDG to the PCR or RT-PCR system and treating it at room temperature for 10 minutes can fully eliminate potential contamination from previous PCR amplification products containing dU. This product is not only suitable for PCR, but also for qPCR, RT-PCR, qRT-PCR and other systems.

Source	Recombinant expression in E. coli
Appearance	Sterile liquid
Storage Buffer	20mM Tris-HCl (pH 7.5), 50mM NaCl, 0.1mM EDTA, 1mM DTT, 50% (v/v) glycerol
Enzyme Concentration	1U/μL
Purity	Free of endonuclease, exonuclease, RNase, and phosphatase activities other than UDG enzyme activity
Activity Definition	One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. Activity is measured by release of [³H]-uracil in a 50 µl Standard Taq Reaction Buffer containing 0.2 µg DNA (10⁴–10⁵ cpm/µg) in 30 minutes at 37℃.

Components and Description

H292766	Component	100U	5×100U	Storage
H292766A	Heat-labile Bacterial UDG (1U/μl)	100µl	5×100µl	-20℃. Avoid freeze/thaw cycle.
H292766B	10× Heat-labile Bacterial UDG Buffer	500µl	5×500µl	-20℃. Avoid freeze/thaw cycle.

Product Applications
Removal of uracil bases from single-stranded or double-stranded DNA; removal of aerosol contamination from PCR products containing dU; in next-generation sequencing (NGS) applications, mainly used for the construction of mRNA-directed sequencing libraries; used to improve the efficiency of site-directed mutagenesis methods and obtain highly labeled oligonucleotide probes; used in PCR, qPCR, RT-PCR, RT-qPCR systems.
Product Advantages
This product is heat-labile and can be rapidly and irreversibly inactivated by incubation at 50℃ for 10 minutes.

Instructions for Use
Setup of PCR reaction system:
a. Set up the PCR or RT-PCR reaction system according to the conventional reaction system, and add Heat-labile Bacterial UDG to a final concentration of 0.02U/μl, then mix well. Usually, only the PCR or RT-PCR buffer needs to be added, and there is no need to add UDG buffer.
b. Incubate at 25℃ for 5-10 minutes (to remove potential contamination from PCR amplification products containing dUTP), then proceed to the PCR or RT-PCR program.
Precautions
(1) Heat-labile Bacterial UDG enzyme is active in most PCR or RT-PCR systems. However, for self-used PCR or RT-PCR systems, it is recommended to test the compatibility with the used system for the first time. Usually, take the PCR amplification product containing dUTP, add an appropriate amount of UDG with reference to Figure 1, and observe whether the PCR amplification product containing dUTP can be effectively degraded. This product is inhibited by high ionic strength (>100mM).

(2) For dNTP/dUTP, it is recommended to purchase D745378 dNTP/dUTP Mixture (2.5mM each/5mM) from Aladdin.

(3) The activity of Heat-labile Bacterial UDG enzyme on double-stranded DNA is lower than that on single-stranded DNA. It has activity on small U-DNA oligonucleotides and dUMP, but no activity on RNA or normal DNA without dUTP.

(4) The activity of Heat-labile Bacterial UDG enzyme is not dependent on metal ions, and it has activity in the presence of Mg²⁺ or EDTA.

(5) The abasic sites of the DNA strand generated by Heat-labile Bacterial UDG enzyme digestion can be removed by heating, alkali treatment, or endonuclease treatment. Usually, the heating step during the PCR reaction can ensure that the sites digested by UDG enzyme are completely cleaved.

(6) Heat-labile Bacterial UDG enzyme can remove accidentally contaminated PCR products containing dUTP before the PCR reaction, thereby avoiding false positive PCR results caused by contamination.

(7) If Heat-labile Bacterial UDG enzyme needs to be applied to One-Step RT-PCR or One-Step qRT-PCR systems, a thermostable reverse transcriptase should be selected, and the reverse transcription temperature needs to be set to 50-55℃.

(8) This product is only for scientific research by professionals, and shall not be used for clinical diagnosis or treatment, nor for food or drugs. It shall not be stored in ordinary residences.

(9) For your safety and health, please wear a lab coat and disposable gloves during operation.

生物活性

1 U/μL

表达系统

E. coli

种属

无载体

Yes

无动物源

Yes

来源

重组表达

储存与运输

物理形态	液体
储存缓冲液	20mM Tris-HCl, 50mM KCl, 0.1mM EDTA, 1mM DTT, 50%(v/v) Glycerol, pH7.5.
浓度	1 U/μL
储存温度	-20°C储存,避免反复冻融
运输条件	超低温冰袋运输
稳定性与储存	长期储存-20℃；收货后建议分装，避免反复冻融。
CAS编号和信息	59088-21-0
酶学委员会编号	EC 3.2.2.27
单位定义	One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. Activity is measured by release of [3H]-uracil in a 50 µl Standard Taq Reaction Buffer containing 0.2 µg DN
分子类型	酶

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引用文献

1. Ruyue Cong, Kaiyue Hu, Dandan Hu, Jingran Sun, Jinglu Yao, Zhiyong Guo, Sui Wang, Yufang Hu. (2025) Ag-enhanced MOF-based electrochemical biosensor for sensitive detection of uracil-DNA glycosylase activity in cellular systems. MICROCHEMICAL JOURNAL, 212 (113236). [10.1016/j.microc.2025.113236]

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