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Related Concept Videos

RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...

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Related Experiment Video

Updated: Jun 30, 2026

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
07:10

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Published on: July 8, 2025

Amplification-Free Detection of Highly Structured RNA Molecules Using SCas12aV2.

Teng Hu1, Youyang Pei1, Zhaoyi Hu1

  • 1Hubei Light Industry Technology Institute, Hubei, China.

Bio-Protocol
|June 29, 2026
PubMed
Summary
This summary is machine-generated.

The new SCas12aV2 assay enables direct RNA detection without amplification, overcoming limitations of previous CRISPR/Cas12a methods for complex RNA molecules and single-nucleotide polymorphism analysis.

Keywords:
CRISPR/Cas12aSCas12a systemScaffold RNASpacer RNAcrRNA

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Published on: December 23, 2022

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • CRISPR/Cas12a systems offer powerful molecular diagnostics but often require pre-amplification for RNA detection.
  • Existing SCas12a assays detect miRNA without amplification but struggle with long, structured RNA.
  • Accurate quantification of diverse RNA targets remains a challenge in molecular diagnostics.

Purpose of the Study:

  • To develop an enhanced CRISPR/Cas12a system (SCas12aV2) for direct RNA detection.
  • To enable amplification-free quantification of long RNA molecules with complex secondary structures.
  • To improve specificity for single-nucleotide polymorphism (SNP) detection.

Main Methods:

  • Development of the SCas12aV2 system utilizing a split-crRNA approach with Cas12a.
  • Preparation of the SCas12a system for direct RNA target detection.
  • Application of SCas12aV2 for detecting RNA from clinical samples and distinguishing PAM-distal sites.

Main Results:

  • SCas12aV2 efficiently detects long-chain RNA molecules with complex secondary structures.
  • The system demonstrates high specificity in single-nucleotide polymorphism (SNP) applications.
  • The entire experimental procedure is completed in under one hour.

Conclusions:

  • SCas12aV2 advances CRISPR-based diagnostics by enabling direct, amplification-free RNA detection.
  • The enhanced system broadens the scope of Cas12a applications to complex RNA and SNP analysis.
  • SCas12aV2 offers a rapid and specific diagnostic tool for clinical RNA targets.