A microRNA detection system based on padlock probes and rolling circle amplification.
The differential expression and the regulatory roles of microRNAs (miRNAs) are being studied intensively these years. Their minute size of only 19-24 nucleotides and strong sequence similarity among related species call for enhanced methods for reliable detection and quantification. Moreover, miRNA expression is generally restricted to a limited number of specific cells within an organism and therefore requires highly sensitive detection methods. Here we present a simple and reliable miRNA detection protocol based on padlock probes and rolling circle amplification. It can be performed without specialized equipment and is capable of measuring the content of specific miRNAs in a few nanograms of total RNA.
Jonstrup
Søren Peter
SP
Department of Molecular Biology, University of Aarhus, C.F. Møllers Allé, DK-8000 Arhus C, Denmark.
Koch
Jørn
J
Kjems
Jørgen
J
D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D011088:DNA Ligases / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009447:Neuroblastoma / Q000473:pathology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000235:genetics*; D015347:RNA Probes / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics*; D000079962:RNA, Circular; D013698:Templates, Genetic
D035683:MicroRNAs; D015347:RNA Probes; D000079962:RNA, Circular; D012313:RNA; D004259:DNA-Directed DNA Polymerase; D011088:DNA Ligases