Next-generation sequencing has opened up new avenues for the identification of microRNAs (miRNAs) and their corresponding roles in abiotic and biotic stress responses. Recently, a plethora of evidence suggests a canonical action of miRNA-mRNA interactions to regulate plant systems biology at the posttranscriptional level, thus leading to the gain or loss of genetic adaptation in plants. In this chapter, we present a detailed protocol for the identification of miRNA targets using six different prediction tools.