CRISPR-Cas12a based internal negative control for nonspecific products of exponential rolling circle amplification.

ABSTRACT: False-positive results cause a major problem in nucleic acid amplification, and require external blank/negative controls for every test. However, external controls usually have a simpler and lower background compared to the test sample, resulting in underestimation of false-positive risks. Internal negative controls, performed simultaneously with amplification to monitor the background level in real-time, are therefore appealing in both research and clinic. Herein, we describe a nonspecific product-activated single-stranded DNA-cutting approach based on CRISPR (clustered regularly interspaced short palindromic repeats) Cas12a (Cpf1) nuclease. The proposed approach, termed Cas12a-based internal referential indicator (CIRI), can indicate the onset of nonspecific amplification in an exponenti

SUBMITTER: Tian B 

PROVIDER: S-EPMC7049689 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature