Light-start DNA amplification using light-controlled DNA polymerase

Abstract

Non-specific amplification caused by the uncontrollable activity of natural DNA polymerase greatly limits the application of DNA amplification techniques. Modification of DNA polymerase using hot-start technology is an effective way to improve the accuracy of nucleic acid detection. However, hot-start technology was only suitable for DNA polymerases working at high temperatures. Accordingly, there is an urgent need for more applicable methods of controlling DNA polymerase activity that do not rely on changes in temperature. In this work, we therefore developed a simple and novel approach to construct light-controlled DNA polymerases. These DNA polymerases are efficiently blocked by DNA aptamers and rapidly activated by UV light irradiation. Using the light-controlled DNA polymerases, we successfully developed light-start loop-mediated isothermal amplification (LAMP), light-start PCR, and light-start rolling circle amplification (RCA). Compared with hot-start technology, light-start technology improves the accuracy of DNA amplification reaction over a wider range of temperature conditions, which will further expand the application scenarios of DNA amplification techniques.