A simple and fast method for profiling microRNA expression from low-input total RNA by microarray

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). 2012 IUBMB IUBMB Life, 64(7): 612616, 2012