Taqman Probe Synthesis Services
As a kind of oligonucleotide, Taqman fluorescent probe is labeled with a fluorescent reporter group and a fluorescent quenching group at both ends. When the probe is intact, the fluorescence signal from the reporting group can be absorbed by the quenching group, and the fluorescence monitoring system cannot detect the fluorescence signal. During PCR amplification process, the probe was digested and degraded by the 5-3 exonuclide activity of Taq enzyme, and the fluorescence report group and the fluorescence quenching group were separated. So, every time a DNA strand is amplified, a fluorescent molecule is formed. The fluorescence monitoring system by real-time detection of the fluorescent signal to realizes the accumulation of fluorescent signal and the formation of PCR products.
Fluorescence signal generation mechanism of TaqMan probe
Creative Biolabs is a professional biotechnology services platform providing outsourced oligonucleotide synthesis services. We have professional researchers and strict quality control to ensure the synthesis of locked TaqMan probe.
TaqMan probes are divided into two types according to the fluorescence quenching group labeled at the 3'end:
- Ordinary TaqMan probes
- TaqMan MGB probes
TaqMan MGB probe
Design principles of TaqMan probe:
- Conservation: The probe must be absolutely conservative, and sometimes the typing is determined by the probe alone.
- Probe length: The length of the Taqman probe is preferably between 25-32 bp, and the Tm value is between 68-72°C, to ensure that the Tm value of the probe is 10°C higher than the Tm value of the primer.
- The tag name of the probe: the position and length of the probe in the genome should be marked.
- Probe Tm value calculation: Calculate the Tm value to ensure that the GC content in the probe is 30-80%, and avoid multiple repeated bases in the probe.
- Evaluation of the probe: In theory, choose the higher dG value.
- Avoid G on the 5'end: When a single G base is connected to the FAM fluorescent reporter group, G can quench the fluorescent signal emitted by the FAM group, resulting in false negatives.
- The position between the Taqman probe and the primer: Taqman probe should be close to the upstream primer, that is, the Taqman probe should be close to the upstream primer on the same chain
Advantages of TaqMan probe method:
- High specificity
- Good repeatability
- Suitable for specific goals
Applicable scope of TaqMan probe method fluorescence quantitative PCR:
- Pathogen detection
- Study disease resistance genes
- Drug efficacy assessment
- Diagnosis of genetic diseases
- Gene quantitative analysis
- Genetic mutation (SNP) detection
- Genetic diagnosis and individualized drug analysis
References
- Laura Orschler, Shelesh Agrawal, et al. Lost in translation: the quest for Nitrosomonas cluster 7‐specific amoA primers and TaqMan probes. Microbial Biotechnology. 2020, 6(13): 2069-2076.
- Wang Hai-Hua, Yin Can, et al. Development of a Real-Time TaqMan PCR Method for Absolute Quantification of the Biocontrol Agent Esteya vermicola. Plant Disease. 2020, 6(104): 1694-1700.
*For Research Use Only. Not for use in diagnostic procedures.