化学
清脆的
放大器
DNA
劈开
计算生物学
脱氧核酶
劈理(地质)
分子生物学
纳米技术
聚合酶链反应
基因
生物化学
生物
古生物学
断裂(地质)
材料科学
作者
Cheng Qian,Rui Wang,Hui Wu,Fang Zhang,Jian Wu,Liu Wang
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2019-08-12
卷期号:91 (17): 11362-11366
被引量:93
标识
DOI:10.1021/acs.analchem.9b02554
摘要
The CRISPR/Cas12a (cpf1) system was reported to indiscriminately cleave single-stranded DNA after binding with target DNA strands. This usually required the target DNA strands to contain the protospacer-adjacent motif (PAM) sequence of TTTN. Herein, we found Cas12a can also recognize another PAM sequence of UUUN resulting in activation of its ssDNA collateral cleavage effect. To make this finding useful, by combining with LAMP, we first realized CRISPR/Cas12a for directly visualized DNA detection at the single-copy level. By treating with UDG enzyme, we made this system free from residual amplicon contamination, which is a big problem in this field. Thus, an ultrasensitive and anticontaminant DNA detection platform, namely, UDG and LAMP and CRISPR (ULC). This new finding would help us better understand the mechanism of Cas12a and expand its application.
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