miRNA检测最快最简单的非同位素最新方法.pdf

This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues. Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. In most cases authors are permitted to post their version of the article (e.g. in Word or Tex form) to their personal website or institutional repository. Authors requiring further information regarding Elsevier’s archiving and manuscript policies are encouraged to visit: /copyright Authors personal copy Detecting miRNAs by liquid hybridization and color development Xiangqi Li a,b,?, Minjie Ni a, Yonglian Zhang a,b,? a Shanghai 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 b Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China a r t i c l e i n f o Article history: Available online 3 August 2012 Communicated byGray Brewer Keywords: Method MicroRNA Liquid hybridization Avidin–biotin detection system Color development a b s t r a c t Currently, two methods, PCR and Northern blot, are widely used to detect individual microRNAs (miRNA). Although PCR is highly sensitive, false positives and difficulties of primer design discourage its use. While a Northern blot is an effective tool, traditional Northern blot protocols are complicated, time-consuming, and usually inconvenient for users. Liquid Northern blot methods are rapid but require instruments for detection of fluorescent signals. Here, we describe an alternative protocol, liquid hybridization and color development (LHCD), based on the rapidity of liquid hybridization and the signal amplification of avidin– biotin complex (ABC) for detection. LHCD can di