title: A microRNA detection system based on padlock probes and rolling circle amplification.
abstract: 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.
authors.affiliation: Department of Molecular Biology, University of Aarhus, C.F. Møllers Allé, DK-8000 Arhus C, Denmark.
authors.forename: Søren Peter
authors.initials: SP
authors.lastname: Jonstrup
authors.forename: Jørn
authors.initials: J
authors.lastname: Koch
authors.forename: Jørgen
authors.initials: J
authors.lastname: Kjems
mesh_terms: 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
chemical_list: D035683:MicroRNAs; D015347:RNA Probes; D000079962:RNA, Circular; D012313:RNA; D004259:DNA-Directed DNA Polymerase; D011088:DNA Ligases