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Cas12a-based one-pot SNP detection with high accuracy

Cell Insight. 2023-02; 
Hong-Xia Zhang, Caixiang Zhang, Shuhan Lu, Xiaohan Tong, Kun Zhang, Hao Yin, Ying Zhang
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Proteins, Expression, Isolation and Analysis … Similarly, the recombinant His-TrSAM protein was purified using High Affinity Ni-NTA Resin (GenScript, Nanjing, China) from E.coli under identical condition. A weight of 20 μg purified … Get A Quote

摘要

CRISPR-Cas12a based one-pot detection system has been used in nucleic acid detection and diagnosis. However, it is not sensitive enough to distinguish single nucleotide polymorphisms (SNP), which has greatly restricted its application. To overcome these limitations, we engineered a LbCas12a variant with enhanced sensitivity against SNP, named seCas12a (sensitive Cas12a). SeCas12a-based one-pot SNP detection system is a versatile platform that could use both canonical and non-canonical PAM, and was almost not limited by mutation types to distinguish SNPs located between position 1 to 17. The use of truncated crRNA further improved SNP specificity of seCas12a. Mechanistically, we found only when the -cleavage was... More

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