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SHERLOCK: nucleic acid detection with CRISPR nucleases.

Nat Protoc. 2019; 
Kellner Max J,Koob Jeremy G,Gootenberg Jonathan S,Abudayyeh Omar O,Zhang
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Proteins, Expression, Isolation and Analysis (Optional, needed only if not using eStain L1 Protein Staining System) Destain solution (GenScript, cat. no. L00659) Get A Quote

摘要

Rapid detection of nucleic acids is integral to applications in clinical diagnostics and biotechnology. We have recently established a CRISPR-based diagnostic platform that combines nucleic acid pre-amplification with CRISPR-Cas enzymology for specific recognition of desired DNA or RNA sequences. This platform, termed specific high-sensitivity enzymatic reporter unlocking (SHERLOCK), allows multiplexed, portable, and ultra-sensitive detection of RNA or DNA from clinically relevant samples. Here, we provide step-by-step instructions for setting up SHERLOCK assays with recombinase-mediated polymerase pre-amplification of DNA or RNA and subsequent Cas13- or Cas12-mediated detection via fluorescence and c... More

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