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Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Profiling human gene expression with cDNA microarrays.

Y Jiang¹, J Lueders, A Glatfelter

¹National Human Genome Research Institute, NIH, Bethesda, Maryland, USA.

Current Protocols in Human Genetics

|April 23, 2008

Summary

Genomics research provides 3-end complementary DNA (cDNA) clones for gene expression analysis. This study details methods for using these clones in high-throughput gene expression assays via cDNA microarrays.

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Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Genomics initiatives have increased the availability of complementary DNA (cDNA) clones representing the 3 ends of human genes.
These clones are valuable resources for studying gene expression patterns.

Purpose of the Study:

To describe methods for utilizing 3-end cDNA clones in gene expression analysis.
To enable a highly parallel assay for monitoring gene expression levels.

Main Methods:

Preparation of complementary DNA (cDNA) microarrays.
Extraction of RNA from specific cell types.
Labeling of RNA with fluorescent markers to create cDNA representations.
Hybridization of labeled cDNA to microarrays for detection.

Main Results:

Established protocols for cDNA microarray preparation.
Developed methods for RNA extraction and fluorescent labeling.
Demonstrated successful hybridization of labeled cDNA to microarrays, enabling gene expression detection.

Conclusions:

The described methods allow for the effective use of 3-end cDNA clones in high-throughput gene expression profiling.
This approach facilitates parallel analysis of gene expression using readily available genomics resources.