Benzonase-Based Closed-Tube DNA Degradation Strategy for Elimination of Nucleic Acid Contamination

Ineffective contamination prevention strategies significantly limit the deployment of nucleic acid testing in point-of-care (POC) settings. We herein propose a benzonase-based closed-tube strategy that degrades amplification products of nucleic acid. During the amplification reaction in the tube, Benzonase is kept on the inner surface of the tube cap. The subsequent shaking allows efficient degradation of the amplification products. Experimental results demonstrate that Benzonase remains active after high-temperature treatment, which does not interfere with real-time signal detection. We have further evaluated its ability to reduce carry-over contamination, and a significant decrease in contamination events can be observed. Moreover, this strategy is compatible with POC techniques, such as colorimetric and electrochemical assays. It could be integrated with various nucleic acid amplification strategies and offers the potential to support a wider range of signal output formats.