Characterization and application of aptamers for Taq DNA polymerase selected using an evolution-mimicking algorithm

Area of Science:

• Molecular Biology
• Biotechnology
• Biochemistry

Background:

• DNA aptamers are nucleic acid-based ligands with high specificity and affinity.
• Taq DNA polymerase and Tth DNA polymerase are crucial enzymes in molecular biology, particularly for PCR.
• Inhibiting DNA polymerase activity can improve PCR specificity and reduce non-specific amplification.

Purpose of the Study:

• To further improve the inhibitory activity of DNA aptamers against Taq DNA polymerase.
• To characterize the most potent DNA aptamers identified.
• To investigate the applicability of these aptamers in inhibiting Tth DNA polymerase and modulating PCR specificity.

Main Methods:

• Evolution-mimicking algorithm (EMA) for aptamer identification and optimization.
• In vitro characterization of aptamer inhibitory activity against DNA polymerases.
• Assessment of aptamer performance in polymerase chain reaction (PCR) for low-copy-number gene detection.

Main Results:

• Identified DNA aptamers with enhanced inhibitory effects on Taq DNA polymerase.
• The most potent aptamer demonstrated inhibitory activity against both Taq DNA polymerase and Tth DNA polymerase.
• Application of the aptamer in PCR significantly improved the specificity of detecting a low-copy-number target gene.

Conclusions:

• Optimized DNA aptamers effectively inhibit Taq and Tth DNA polymerases.
• These aptamers represent a valuable tool for enhancing PCR specificity and sensitivity.
• The findings support the use of aptamers for precise molecular diagnostics and research applications.