Quantification of small non-coding RNAs allows an accurate comparison of miRNA expression profiles.
MicroRNAs (miRNAs) are highly conserved approximately 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.
Masotti
Andrea
A
Gene Expression and Microarrays Laboratory, Bambino Gesú Children's Hospital, P. za S. Onofrio 4, 00165 Rome, Italy. andrea.masotti@opbg.net
Caputo
Viviana
V
Da Sacco
Letizia
L
Pizzuti
Antonio
A
Dallapiccola
Bruno
B
Bottazzo
Gian Franco
GF
D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D016018:Least-Squares Analysis; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D035683:MicroRNAs; D012313:RNA