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Updated: Jul 16, 2026
07:27
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
Direct and sensitive miRNA profiling from low-input total RNA
Hui Wang1, Robert A Ach, Bo Curry
1Agilent Technologies, Inc., Agilent Laboratories, Santa Clara, California 95051, USA. hui_wang@agilent.com
RNA (New York, N.Y.)
|November 16, 2006
Summary
Researchers developed a sensitive microRNA (miRNA) profiling assay using novel microarray probes for accurate detection. This method enables precise quantification of miRNA content in various samples, including clinical tissues.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- MicroRNA (miRNA) profiling is crucial for understanding gene regulation and disease.
- Existing methods may lack sensitivity, specificity, or require complex protocols.
- Accurate and efficient miRNA detection is needed for research and clinical applications.
Purpose of the Study:
- To develop a sensitive, accurate, and multiplexed assay for microRNA profiling.
- To enable precise quantification of miRNA content in biological samples.
- To create a versatile assay applicable to various sample types, including clinical specimens.
Main Methods:
- Development of a novel microarray probe design with sequence and size discrimination capabilities.
- Implementation of a highly efficient, single-vial labeling protocol for total RNA (120 ng) using Cy3 or Cy5.
- Direct labeling without fractionation or amplification, ensuring simplicity and speed.
Main Results:
- Achieved highly specific detection of closely related mature miRNAs.
- Established a precise and accurate linear dynamic range from 0.2 amol to 2 fmol of input miRNA.
- Demonstrated suitability for formalin-fixed paraffin-embedded clinical samples.
Conclusions:
- The developed assay provides sensitive and accurate miRNA profiling.
- The method allows for quantitative estimation of miRNA content in studied tissues.
- The assay supports rapid probe design for simultaneous measurement of known and novel human miRNAs.
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