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Argonaute-mediated system for supersensitive and multiplexed detection of rare mutations

biorxiv. 2020; 
Qian Liu,  Xiang Guo,  Guanhua Xun,  Zhonglei Li,  Yuesheng Chong,  Litao Yang,  Hongxia Wang,  Fengchun Zhang,  Shukun Luo,  Zixin Deng,  Kai Li,  Yan Feng
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摘要

The ability to detect rare mutations has revolutionized diagnosis and monitoring of tumors, but is limited by the shortage of sensitive, cost-effective and high coverage methods for identification of extremely low abundant mutations. Here, we establish a single-tube multiplex PCR system by employing thermophilic Argonaute-derived DNA-guided nuclease for highly efficient rare mutation detection, referred to as A-Star (Argonaute-directed specific target enrichment and detection), that combines the selective cleavage of the wild type DNA in the DNA denaturation step and the followed amplification of mutant DNA during PCR. A-Star enables easy detection and quantitation of rare mutations originally as low as 0.01% i... More

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