title: A simple and fast method for profiling microRNA expression from low-input total RNA by microarray.
abstract: Analysis of mature microRNA (miRNA) expression is important to understand its physiological functions and pathological implications. Microarray is a powerful technology to profile global miRNA expression. In this study, we developed a rapid miRNA labeling method by which miRNA was directly labeled in total RNA for microarray detection. This method consists of RNA tailing by poly(A) polymerase, reverse transcription, and template-switching by moloney murine leukemia virus (MMLV) reverse transcriptase. After these reactions, the small RNA cDNA was ready for labeling for microarray detection. The whole process of prearray sample preparation was dramatically shortened to 2 h. Furthermore, this method allows very limited starting total RNA (100 ng) for microarray analysis. Our data showed that the results from our method were highly consistent with that of real-time polymerase chain reaction (PCR).
authors.affiliation: School of Life Sciences, Shanghai University, Shanghai 200444, China. zhaobotao@shu.edu.cn
authors.forename: Botao
authors.initials: B
authors.lastname: Zhao
authors.forename: Li
authors.initials: L
authors.lastname: Jin
authors.forename: Jiali
authors.initials: J
authors.lastname: Wei
authors.forename: Zhongliang
authors.initials: Z
authors.lastname: Ma
authors.forename: Weijun
authors.initials: W
authors.lastname: Jiang
authors.forename: Lijun
authors.initials: L
authors.lastname: Ma
authors.forename: Youxin
authors.initials: Y
authors.lastname: Jin
mesh_terms: D000818:Animals; D018076:DNA, Complementary / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D046228:Microarray Analysis / Q000379:methods*; D008979:Moloney murine leukemia virus / Q000235:genetics; D016133:Polymerase Chain Reaction; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000378:metabolism; D013997:Time Factors
chemical_list: D018076:DNA, Complementary; D035683:MicroRNAs; D012313:RNA; D011062:Polynucleotide Adenylyltransferase