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Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays
Published on: December 3, 2010
Measuring microRNAs: comparisons of microarray and quantitative PCR measurements, and of different total RNA prep
Robert A Ach1, Hui Wang, Bo Curry
1Agilent Laboratories, Agilent Technologies, Santa Clara, CA 95051, USA. robert_ach@agilent.com
BMC Biotechnology
|September 12, 2008
Summary
MicroRNA (miRNA) expression analysis using microarrays and quantitative RT-PCR (qPCR) generally show excellent correlation. However, researchers should be aware of potential discrepancies in miRNA measurements due to different sample preparation methods.
Area of Science:
- Molecular Biology
- Genomics
Background:
- MicroRNAs (miRNAs) are crucial regulators of cellular processes.
- Accurate measurement of miRNA expression is vital for biological research.
- Microarrays and quantitative RT-PCR (qPCR) are common methods for miRNA quantification.
Purpose of the Study:
- To compare the correlation of miRNA expression levels measured by microarrays and qPCR.
- To assess the impact of different total RNA preparation methods on miRNA measurements.
Main Methods:
- Agilent miRNA microarrays were used to measure 470 human miRNAs in nine tissues.
- Quantitative RT-PCR (qPCR) was employed to profile 61 miRNAs in the same tissues.
- Three common total RNA sample preparation methods were evaluated using miRNA microarrays.
Main Results:
- A high correlation (R > 0.9) was observed for most miRNAs (53/60) between microarray and qPCR methods.
- Differences in low-correlation miRNAs were not due to assay sensitivity.
- Most total RNA preparation methods yielded comparable miRNA levels, with a few exceptions noted.
Conclusions:
- Microarray and qPCR methods demonstrate excellent correlation for miRNA expression analysis.
- Different total RNA preparation methods generally yield consistent results, but variations exist for specific miRNAs.
- Researchers must consider potential variations when comparing miRNA data across different analytical or sample preparation techniques.
