Smartseq
SMART-seq, short for Switching Mechanism at the 5' end of RNA Transcript, is a method for generating full-length cDNA libraries from small amounts of RNA, notably from single cells. It enables transcriptional profiling with near full-length transcripts, facilitating detection of splice isoforms and other transcript features. The protocol begins with reverse transcription using an oligo-dT primer to capture mRNA, followed by template-switching at the 5' end of the cDNA to add a universal priming site. This enables subsequent PCR amplification of the full-length cDNAs, which are then prepared into sequencing libraries compatible with high-throughput platforms such as Illumina.
Variants of SMART-seq include SMART-seq2, an improved version that increases sensitivity and uniformity of transcript coverage,
Applications of SMART-seq and its variants span single-cell transcriptomics, developmental biology, neuroscience, and cancer research, where