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

Abstract

SummaryAnalysis 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). IUBMBIUBMB Life, 64(7): [612][613][614][615][616] 2012

Ultrasensitive optical biosensor for detection of miRNA-155 using positively charged Au nanoparticles

“…Therefore, overexpression of miR-155 suggests a risk factor for breast cancer 10 . Currently, methods for detection of miRNA mainly include reverse transcription polymerase chain reaction (RT-PCR) 11 – 13 , northern blotting 14 , 15 , microarray technique 16 , 17 , in situ hybridization 18 – 20 and so on. Despite the advantages, these methods still suffer some disadvantages such as requiring expensive instruments and reagents, miRNA labeling, large amounts of miRNA and relatively pure miRNA samples 21 .…”