The phosphorylation modification of primers and probes in molecular biology techniques confers additional functionalities and applications. Here are the effects of phosphorylation modification on primers and probes:

1. Enhanced Stability: Phosphorylation modification enhances the stability of primers and probes, making them more durable under experimental conditions. This is particularly crucial for applications requiring prolonged PCR amplification, hybridization experiments, and other molecular biology techniques with high stability demands.
2. Increased Affinity: Phosphorylation modification increases the affinity of primers and probes to target sequences, thereby enhancing the sensitivity and specificity of PCR amplification or hybridization experiments. This is particularly important for applications detecting low-abundance targets or specific sequences.
3. Regulation of Primer Specificity: In PCR amplification, phosphorylation modification can regulate the specificity of primers, reducing nonspecific amplification or primer dimer formation. This helps improve the purity and specificity of PCR products.
4. Enhanced Probe Stability and Specificity: Phosphorylation modification enhances the stability and specificity of probes binding to target molecules, improving probe performance in hybridization experiments. This is crucial for probe design in techniques such as in situ hybridization, Northern blotting, Southern blotting, etc.
5. Modulation of Signal Amplification: In certain detection techniques like fluorescence PCR or fluorescence in situ hybridization, phosphorylation modification of primers or probes can modulate the signal amplification, thereby enhancing the sensitivity and dynamic range of detection.
6. Improvement of Inhibition Effects: Under specific experimental conditions, such as complex sample matrices or environments containing RNAases, phosphorylation modification of primers and probes can reduce inhibitory effects, increasing the success rate of experiments.

Overall, phosphorylation modification of primers and probes provides molecular biology techniques with greater design flexibility and application potential, improving stability, specificity, and sensitivity of experiments, thereby facilitating target analysis and detection more effectively.

GenCefe provides customized DNA oligo modification/labeling services, including more than 100 modifications such as biotin, digoxigenin, phosphate, thiol, sulfhydryl, amino, special bases, and a variety of fluorescent groups. 100% of the delivered modified probes/primers have passed mass spectrometry detection to ensure accuracy and purity, meeting the needs of scientific research and industrial-grade modified primers/probes.