Small RNA sequencing is crucial for studying short RNA molecules (18-30 nt) involved in gene regulation. It analyzes miRNAs and small RNAs, shedding light on gene regulation, development, and disease. The process involves RNA extraction, size selection, adapter ligation, library amplification, size selection, sequencing, and bioinformatics analysis to identify and quantify small RNAs. High-throughput sequencing reveals small RNA profiles, which are analyzed through bioinformatics to identify miRNAs, siRNAs, and piRNAs, aiding in understanding their roles in cellular processes.
Libraries are prepared using the SMARTer smRNA-Seq Kit (Takara) that is compatible with total RNA or enriched small RNA, incorporating SMART technology and primers with locked nucleic acids (LNAs). Users can analyze various small RNA species and create complex sequencing libraries. Illumina adapters and index sequences are added during library amplification, ensuring unbiased representation of diverse small RNA species.