Temperature Sensitive UNG

Temperature Sensitive UNG (TS-UNG) is a obtained by recombinant expression in E. coli. The enzyme catalyzes the release of free uracil from uracil-containing single- and double-stranded DNA and is inactive against RNA. Compared with the conventional UNG enzyme of E. coli gene origin, TS-UNG enzyme has higher activity at low temperatures (20℃~37℃) and is temperature sensitive and easily inactivated (50℃), avoiding the degradation of dUTP-containing amplification products at room temperature by the residual activity that may remain after inactivation of the conventional UNG enzyme. Therefore, TS-UNG enzyme is not only suitable for PCR contamination prevention reaction, but also matches well with the RT-PCR amplification program and can be used in RT-PCR contamination prevention reaction.

Recommended Application

Contamination Prevention Amplification

Storage Condition

-20°C for long term storage, should be mixed well before use, avoid frequent freeze-thaw.

Storage buffer

20 mM Tris-HCl (pH 7.5) , 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, Stabilizer, 50% Glycerol.

Unit Definition

The amount of enzyme required to degrade 1µg of single-stranded DNA containing dU bases in 1 hour at 37°C is 1 unit of activity (U).

Quality Control

1.SDS-PAGE electrophoretic purity greater than 98%

2.Degradation activity, batch-to-batch control, stability

3.No exogenous nuclease activity , no exogenous endonuclease or exonuclease contamination.

Instructions

Note: a: If used for qPCR/qRT-PCR, the fluorescent probe should be added into the reaction system. Usually, a final primer concentration of 0.2 μM can give good results; when the reaction performance is poor, the primer concentration can be adjusted in the range of 0.2-1 μM. Usually, the probe concentration is optimized in the range of 0.1-0.3 μM. Concentration gradient experiments can be performed to find the best combination of primer and probe.

Notes

1.The optimal reaction temperature of TS-UNG enzyme is relatively low, and it can be optimized in the range of 20℃~37℃, the dosage of enzyme and the reaction time can be optimized in the range of 0.1~0.5 U, 5~10 min; and the enzyme can be inactivated in the process of reverse transcription.

2.It is suitable for PCR and RT-PCR to prevent contamination.

3.Avoid frequent freeze-thaw, and do not expose to large temperature fluctuations.

4.Different genes to be amplified have different utilization efficiency of dUTP and sensitivity to UNG enzyme, therefore, if the use of UNG system leads to a decrease in detection sensitivity, the reaction system should be adjusted and optimized, if you need technical support, please contact our company.