本公司生产的 Uracil-DNA Glycosylase (Heat-labile, Cod)，也称 Heat-labile Cod UDG，即热敏型鳕鱼 UDG，是来源于大西洋鳕鱼(Gadus morhua cod)，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 Cod UDG 室温处理 5min，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 |
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| 外观 (Appearance) | 无菌液体 | | 保存液 (Storage Buffer) | 50mM Tris-HCl (pH 7.5), 100mM NaCl, 0.5mM EDTA, 1mM DTT, 0.1% Triton X-100, 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 the amount of enzyme required to liberate 1 nmol uracil from dU-containing DNA in one hour at 37℃. |
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组分和说明
| U751666 | Component | 200U | 1KU | 5×1KU | Storage | | U751666A | Heat-labile Cod UDG (1U/μl) | 200µl | 1ml | 5×1ml | -20℃. Avoid freeze/thaw cycle. | | U751666B | 10× Heat-labile Cod UDG Buffer | 1ml | 5ml | 5×5ml | -20℃. Avoid freeze/thaw cycle. |
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产品应用
去除含尿嘧啶的单链或双链 DNA；去除含 dU 的 PCR 产物气溶胶污染；DNA 修复与突变检测；提高 PCR 产物的克隆效率；提高点诱变的效率；研 究蛋白质-DNA 相互作用。用于 PCR、qPCR、RT-PCR、RT-qPCR、one-step RT-PCR 体系。
产品优势
产品不仅适用于 PCR，也适用于 qPCR、RT-PCR 和 qRT-PCR 等体系。
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 Cod UDG 至最终浓度为 0.01U/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 5min (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 Cod UDG 酶在大多数 PCR 反应缓冲液中均具有活性，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，加入适量 UDG，观察能否有效降解含 dUTP 的 PCR 扩增产物。
（2）dNTP/dUTP 推荐选购阿拉丁的 D745378 dNTP/dUTP Mixture (2.5mM each/5mM)。
（3）Heat-labile Cod UDG 长期储存请置于-80℃保存，常规使用请于-20℃保存。-80℃保存会结冻，尽量避免反复冻融。
（4）如果需要将 Heat-labile Cod 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℃。
（5）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。
（6）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, Cod) produced by our company, also known as Heat-labile Cod UDG or thermolabile cod UDG, is derived from Atlantic cod (Gadus morhua). It catalyzes the hydrolysis of the N-glycosidic bond between the uracil (dU) base and deoxyribose in DNA strands 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 its anti-contamination function is as follows: An appropriate amount of dUTP is added to the PCR reaction, and dUTP replaces dTTP to be incorporated into DNA, forming PCR amplification products containing dU bases. During subsequent PCR reactions, Heat-labile Cod UDG is used to selectively cleave the single-stranded or double-stranded DNA containing dU (which may be brought in by contamination from previous PCR amplifications), thereby avoiding the negative impact of possible contamination from previous PCR amplification products on the current PCR amplification.This product is thermolabile and can be rapidly and irreversibly inactivated by incubation at 50°C for 10 minutes. Before PCR or RT-PCR reactions, adding Heat-labile Cod UDG to the PCR or RT-PCR system and incubating at room temperature for 5 minutes can fully eliminate potential contamination from previous PCR amplification products containing dUTP. This product is suitable not only for PCR but also for systems such as qPCR, RT-PCR, and qRT-PCR.
| Source | Recombinant expression in E. coli |
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| Appearance | Sterile liquid | | Storage Buffer | 50mM Tris-HCl (pH 7.5), 100mM NaCl, 0.5mM EDTA, 1mM DTT, 0.1% Triton X-100, 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 the amount of enzyme required to liberate 1 nmol uracil from dU-containing DNA in one hour at 37℃. |
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Components and Description
| U751666 | Component | 200U | 1KU | 5×1KU | Storage | | U751666A | Heat-labile Cod UDG (1U/μl) | 200µl | 1ml | 5×1ml | -20℃. Avoid freeze/thaw cycle. | | U751666B | 10× Heat-labile Cod UDG Buffer | 1ml | 5ml | 5×5ml | -20℃. Avoid freeze/thaw cycle. |
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Product Applications
Removal of uracil-containing single-stranded or double-stranded DNA; Removal of aerosol contamination from dU-containing PCR products; DNA repair and mutation detection; Improvement of PCR product cloning efficiency; Improvement of site-directed mutagenesis efficiency; Study of protein-DNA interactions. Applicable to PCR, qPCR, RT-PCR, RT-qPCR, and one-step RT-PCR systems.
Product Advantages
This product is suitable not only for PCR but also for systems such as qPCR, RT-PCR, and qRT-PCR.
Instructions for Use
Setup of PCR reaction system:
a. Prepare the PCR or RT-PCR reaction system according to the standard protocol, and add Heat-labile Cod UDG to a final concentration of 0.01 U/μl, then mix well. Usually, only the PCR or RT-PCR buffer is required; no additional UDG buffer is needed.
b. Incubate at 25°C for 5 minutes (to remove potential contamination from previous dUTP-containing PCR amplification products), then proceed with the PCR or RT-PCR program.
Precautions
(1) Heat-labile Cod UDG is active in most PCR reaction buffers. However, for self-prepared PCR or RT-PCR systems, it is recommended to test compatibility before first use. Typically, take dUTP-containing PCR amplification products, add an appropriate amount of UDG, and observe whether the dUTP-containing PCR products are effectively degraded.
(2) For dNTP/dUTP, it is recommended to purchase Aladdin’s D745378 dNTP/dUTP Mixture (2.5mM each/5mM).
(3) For long-term storage, store Heat-labile Cod UDG at -80°C; for regular use, store at -20°C. Storage at -80°C will cause freezing, so repeated freeze-thaw cycles should be avoided as much as possible.
(4) If Heat-labile Cod UDG is to be used in One-Step RT-PCR or One-Step qRT-PCR systems, a thermostable reverse transcriptase must be selected, and the reverse transcription temperature should be set to 50°C–55°C.
(5) This product is for scientific research use only by professional personnel. It shall not be used for clinical diagnosis or treatment, nor for food or pharmaceuticals, and shall not be stored in ordinary residential areas.
(6) For your safety and health, wear a lab coat and disposable gloves during operation.
ZJ25F1129674
危险品化学品经营许可证（带存储）