Amplification and sequencing of mature microRNAs in uncharacterized animal models using stem-loop reverse transcription-polymerase chain reaction

Expression of mature microRNA (miRNA) transcripts can be easily measured in many established animal model systems but is difficult to evaluate using conventional methods in new and uncharacterized animal models. In this study, we were able to expand an existing protocol to evaluate miRNA expression in both vertebrate and invertebrate animals for which mature miRNAs remain unsequenced. This method allows the researcher to sequence reverse transcription-polymerase chain reaction products, validating miRNA-specific amplification and providing the opportunity to add to the current body of knowledge of miRNA annotation. (C) 2011 Elsevier Inc. All rights reserved.