title: A versatile method to design stem-loop primer-based quantitative PCR assays for detecting small regulatory RNA molecules.
abstract: Short regulatory RNA-s have been identified as key regulators of gene expression in eukaryotes. They have been involved in the regulation of both physiological and pathological processes such as embryonal development, immunoregulation and cancer. One of their relevant characteristics is their high stability, which makes them excellent candidates for use as biomarkers. Their number is constantly increasing as next generation sequencing methods reveal more and more details of their synthesis. These novel findings aim for new detection methods for the individual short regulatory RNA-s in order to be able to confirm the primary data and characterize newly identified subtypes in different biological conditions. We have developed a flexible method to design RT-qPCR assays that are very sensitive and robust. The newly designed assays were tested extensively in samples from plant, mouse and even human formalin fixed paraffin embedded tissues. Moreover, we have shown that these assays are able to quantify endogenously generated shRNA molecules. The assay design method is freely available for anyone who wishes to use a robust and flexible system for the quantitative analysis of matured regulatory RNA-s.
authors: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical and Health Science Center, Debrecen, Hungary.
authors: Zsolt
authors: Z
authors: Czimmerer
authors: Julianna
authors: J
authors: Hulvely
authors: Zoltan
authors: Z
authors: Simandi
authors: Eva
authors: E
authors: Varallyay
authors: Zoltan
authors: Z
authors: Havelda
authors: Erzsebet
authors: E
authors: Szabo
authors: Attila
authors: A
authors: Varga
authors: Balazs
authors: B
authors: Dezso
authors: Maria
authors: M
authors: Balogh
authors: Attila
authors: A
authors: Horvath
authors: Balint
authors: B
authors: Domokos
authors: Zsolt
authors: Z
authors: Torok
authors: Laszlo
authors: L
authors: Nagy
authors: Balint L
authors: BL
authors: Balint
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D055029:Inverted Repeat Sequences / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D017931:DNA Primers; D034741:RNA, Small Interfering; D038621:Regulatory Sequences, Ribonucleic Acid
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