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Microarray Analysis of Whole-Transcriptome RNAs Including Non-Coding RNAs
Quentin Thuillier1, Isabelle Behm-Ansmant2
1Université de Lorraine, CNRS, IMoPA, Nancy, France.
Methods in Molecular Biology (Clifton, N.J.)
|April 1, 2021
Summary
This study presents a robust method for measuring circulating non-coding RNAs (ncRNAs) in body fluids. The protocol utilizes microarrays for precise quantification, aiding in the development of novel biomarkers for disease detection.
Area of Science:
- Molecular Biology
- Genomics
- Biomarker Discovery
Background:
- Non-coding RNAs (ncRNAs) are crucial in cellular functions and disease pathogenesis.
- Circulating ncRNAs in body fluids show promise as accessible biomarkers.
- Challenges in precise ncRNA measurement include low abundance and population heterogeneity.
Purpose of the Study:
- To describe protocols for extracting and purifying total RNAs from blood samples.
- To detail RNA labeling and hybridization procedures for microarray analysis.
- To enable precise quantification of circulating ncRNAs for biomarker applications.
Main Methods:
- RNA extraction and purification from PAXgene RNA Blood Tubes.
- RNA labeling and hybridization using Affymetrix Clariom™ D microarrays.
- Microarray analysis for expression level and splicing pattern of circulating ncRNAs.
Main Results:
- Established protocols for reliable RNA isolation from blood.
- Demonstrated the utility of microarrays for precise quantification of low-abundance ncRNAs.
- Preserved sample integrity during the analysis process.
Conclusions:
- Microarrays offer a powerful method for analyzing circulating ncRNAs.
- The described protocols facilitate accurate measurement of ncRNA biomarkers.
- This approach supports the development of minimally invasive diagnostic tools.
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