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 |
|---|
| 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℃. |
|
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. |
|
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.
