Article

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

School of Life Sciences, Shanghai University, Shanghai 200444, China.
International Union of Biochemistry and Molecular Biology Life (impact factor: 3.51). 05/2012; 64(7):612-6. DOI:10.1002/iub.1026 pp.612-6
Source: PubMed

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).

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Keywords

labeling
 
mature microRNA
 
microarray analysis
 
MMLV
 
moloney murine leukemia virus
 
pathological implications
 
powerful technology
 
prearray sample preparation
 
profile global miRNA expression
 
rapid miRNA labeling method
 
real-time polymerase chain reaction
 
reverse transcription
 
template-switching
 
total RNA
 
whole process