Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip.

ABSTRACT: Single nucleotide polymorphisms (SNPs) are associated with many human diseases, so accurate and efficient SNP detection is of great significance for early diagnosis and clinical prognosis. This report proposes a universal and high-fidelity genotyping method in microfluidic point-of-care equipment based on the clustered regularly interspaced short palindromic repeat (CRISPR) system. Briefly, by systematically inserting the protospacer-adjacent-motif (PAM) sequence, we improved the universality of the CRISPR/Cas12a based SNP detection; by removing the complementary ssDNA and introducing an additional nucleotide mismatch, we improved the sensitivity and specificity. We preloaded the CRISPR/Cas12a reagents into the point-of-care biochip for automating the process, increasing the stability and

SUBMITTER: Chen Y 

PROVIDER: S-EPMC8179484 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature