Abstract: MicroRNAs ( miRNAs) are a type of endogenous RNAs (~ 22nt) which play an important roles in gene expression and regulation of animals and plants cells by destroying or suppressing targeting RNAs. Thus, the researches related to miRNA have attracted more and more attentions.To quantify a certain miRNA, Real Timequantitative PCR ( RT-qRCR) could be adopted including reverse transcription and PCR amplification using a stem-loop primer designed according to the target miRNA.This method is called SLP-RT-qPCR for short.The miRNA quantification method using SLP-RT-qPCR has a lot of advantages, including high specificity, high sensitivity, wide detection range, precise quantification, etc. Moreover, multiplexing RT- PCR developed in recent years made it possible to determine several kinds of miRNAs simultaneously. All the advantages mentioned above greatly promoted the study of miRNA.In this paper, the development and application of primer design, multi-quantification and fluorescence labeling from the establishment of the method about ten years ago was introduced.The theory and key technologies of this method was also reviewed. The review will be helpful to the application and development of miRNA quantification.