Quantification of Small Non-Coding RNAs Allows an Accurate Comparison of miRNA Expression Profiles

MicroRNAs (miRNAs) are highly conserved ∼22-mer RNA molecules, encoded by plants and animals that regulate the expression of genes binding to the 3′-UTR of specific target mRNAs. The amount of miRNAs in a total RNA sample depends on the recovery efficiency that may be significantly affected by the d...

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Bibliographic Details
Published in:BioMed research international 2009-01, Vol.2009 (2009), p.1-9
Main Authors: Caputo, Viviana, Masotti, Andrea, Da Sacco, Letizia, Pizzuti, Antonio, Dallapiccola, Bruno, Bottazzo, Gian Franco
Format: Article
Language:English
Subjects:
Animals
Apoptosis
Cell Line, Tumor
Cercopithecus aethiops
COS Cells
Electrophoresis, Agar Gel - methods
Electrophoresis, Polyacrylamide Gel - methods
Gene Expression
HeLa Cells
Humans
Identification and classification
Infections
Least-Squares Analysis
Methodology Report
Methods
Microfluidic Analytical Techniques - methods
MicroRNA
MicroRNAs - biosynthesis
MicroRNAs - genetics
MicroRNAs - isolation & purification
Molecular Weight
Polymerase Chain Reaction
RNA - genetics
RNA - isolation & purification
RNA - metabolism
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Summary:MicroRNAs (miRNAs) are highly conserved ∼22-mer RNA molecules, encoded by plants and animals that regulate the expression of genes binding to the 3′-UTR of specific target mRNAs. The amount of miRNAs in a total RNA sample depends on the recovery efficiency that may be significantly affected by the different purification methods employed. Traditional approaches may be inefficient at recovering small RNAs, and common spectrophotometric determination is not adequate to quantify selectively these low molecular weight (LMW) species from total RNA samples. Here, we describe the use of qualitative and quantitative lab-on-a-chip tools for the analysis of these LMW RNA species. Our data emphasize the close correlation of LMW RNAs with the expression levels of some miRNAs. We therefore applied our result to the comparison of some miRNA expression profiles in different tissues. Finally, the methods we used in this paper allowed us to analyze the efficiency of extraction protocols, to study the small (but significant) differences among various preparations and to allow a proper comparison of some miRNA expression profiles in various tissues.
ISSN:1110-7243
2314-6133
1110-7251
2314-6141
DOI:10.1155/2009/659028