title: PolyA RT-PCR-based quantification of microRNA by using universal TaqMan probe.
abstract: Quantification of microRNAs (miRNAs) in tissues under normal and pathological conditions is important for elucidating miRNA functions. Based on a PolyA RT-PCR method we have described (J Zhang et al. Biochem Biophys Res Commun 2008 377:136-140), a modified miRNA quantification method was developed and validated using a universal TaqMan probe complementary to the reverse transcript primer. This method effectively detects miRNA expression in cell lines and tissues. The TaqMan probe is more accurate and reliable than the SYBR Green method since it was free from primer dimers. A series of miRNAs were tested in five different mouse tissues: the method differentiated different miRNAs of the same family. This universal TaqMan probe-based PolyA RT-PCR method showed its advantages in precision, simplicity and high-throughput capability compared with other miRNA-detecting methods.
authors.affiliation: Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, P.O. Box 238, 130 Dongan Rd, Shanghai, 200032, China. luoxin3078@126.com
authors.forename: Xin
authors.initials: X
authors.lastname: Luo
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Huijun
authors.initials: H
authors.lastname: Wang
authors.forename: Yingying
authors.initials: Y
authors.lastname: Du
authors.forename: Lu
authors.initials: L
authors.lastname: Yang
authors.forename: Fengyun
authors.initials: F
authors.lastname: Zheng
authors.forename: Duan
authors.initials: D
authors.lastname: Ma
mesh_terms: D000818:Animals; D051379:Mice; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics*; D011061:Poly A / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D035683:MicroRNAs; D015345:Oligonucleotide Probes; D011061:Poly A