title: One-step real time RT-PCR for detection of microRNAs.
abstract: Rapid and simple methods for microRNA (miRNA) detection are essential for biological research of miRNAs and clinical diagnosis. Here we describe a sensitive and specific real time RT-PCR (also RT-qPCR) method for miRNA quantification. The whole detection process including reverse transcription and PCR is performed in one PCR tube by a one-step operation on a real-time PCR system. The results display a wide linear range from 0.1 amol to 10 fmol with a detection limit of 12.6 zmol for miRNA let-7a detection. Let-7a in small RNA samples extracted from tumor cells has been successfully detected by this method. This method is cost-effective, simple and rapid, and has the advantages in the high-throughput routing assay of given miRNAs, as well as in non-model research that has less specific kits and reagents.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Jingli
authors.initials: J
authors.lastname: Yan
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Cui
authors.initials: C
authors.lastname: Qi
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Liu
authors.forename: Dihua
authors.initials: D
authors.lastname: Shangguan
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers; D006801:Humans; D035683:MicroRNAs / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D035683:MicroRNAs