Small RNA sequencing (sRNA-seq) has greatly transformed the study of molecular biology by allowing thorough analysis of several types of short RNA molecules, such as microRNAs (miRNAs), small interfering RNAs (siRNAs), and piwi-interacting RNAs (piRNAs). This chapter presents a comprehensive methodology for conducting miRNA -seq on biological materials. The text introduces the principles and uses of miRNA-seq, emphasizing its importance in gene regulation, disease processes, and therapeutic interventions. The chapter subsequently explores the essential stages of miRNA-seq, encompassing sample acquisition, RNA isolation, library construction, sequencing, and data interpretation. Every part contains pragmatic suggestions, guidance for resolving issues, and optimal methods to guarantee excellent outcomes. Upon completion of this chapter, readers will possess a comprehensive comprehension of the miRNA-seq workflow and will be adequately prepared to apply this potent approach in their own research endeavors.