A convenient system for highly specific and sensitive detection of miRNA expression
- 1Shanghai Key Laboratory for Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Shanghai 200031, China
- 2Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China
- 3School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China
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↵4 These authors contributed equally to this work.
Abstract
Since the first miRNA was discovered in 1993, miRNAs have become a hotspot for biological research. In order to feed this demand, a robust method is required to detect miRNA gene expression. Development of a detection method is more difficult for miRNAs than for long RNAs, such as mRNA, owing to their small size. Existing methods have limitations; thus, new methods are required. We describe a new system for detecting miRNA expression, which can distinguish miRNA from its precursor and has single-nucleotide resolution. It has single molecule and multiplex detection potential. It may be performed as a polymerase chain reaction (PCR) method, a blotting method, or a macroarray method according to the analyst's preference. This personalized system provides a convenient tool for the detection of miRNA gene expression.
Keywords
Footnotes
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Abbreviations: aLHCD or LH-CD, liquid hybridization and color development with signal amplification; PCR, polymerase chain reaction; RT-PCR, reverse transcription-polymerase chain reaction; Q-PCR, quantitative polymerase chain reaction; ABC, avidin–biotin complex; AP, alkaline phosphatase; BCIP/NBT, 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium; BSA, bovine serum albumin; TBS, Tris-buffered saline; ELISA, enzyme-linked immunosorbent assay
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↵5 Corresponding authors
E-mail lixq{at}sibs.ac.cn
E-mail ylzhang{at}sibs.ac.cn
- Received May 23, 2013.
- Accepted October 20, 2013.
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