SSmRNA + UMIs, the latest improvement in SMART-Seq technology
SMART (Switching Mechanism At 5' end of RNA Template) technology was first described in Biotechniques in 2001 (Zhu et al. 2001) and powers the whole-transcriptome RNA-seq kits in our SMART-Seq portfolio. It leverages the template-switching activity of reverse transcriptases derived from Moloney Murine Leukemia Virus (MMLV) to efficiently capture the 5’ end of transcripts. It is capable of synthesizing full-length cDNA templates up to ~15 kb, incorporating anchor sequences to the ends of the cDNA templates for subsequent cDNA amplification.
Takara Bio introduced the first RNA-seq kit using SMART technology in 2011, and Figure 1 highlights kits in the SMART-Seq portfolio since 2015. These products improved sensitivity while optimizing for specialized applications, such as 3’ differential gene expression, high-throughput analysis, and single-cell RNA-seq specifically formulated for cells with ultra-low RNA content (e.g., peripheral blood mononuclear cells [PBMC], T cells, and B cells). The SMART-Seq mRNA kits have become the industry standard for ultra-low input RNA-seq.
The new SSmRNA + UMIs kit produces final libraries that include 5′-UMI-tagged fragments and internal fragments lacking UMIs, spanning the entire transcript. These libraries enable a combination of full-length RNA-seq with quantitative RNA counting. The complete SMART-Seq chemistry with UMI incorporation is shown in Figure 2.
SSmRNA + UMIs chemistry
SSmRNA + UMIs combined with our free Cogent NGS data analysis tools offer a complete workflow from cDNA synthesis to analysis (Figure 3). Cogent NGS Analysis Pipeline (CogentAP) maps and reports RNA-seq data for full-length UMI-based or UMI-agnostic transcriptome analysis. Included are gene and transcript counting, gene fusion detection, and immune profiling analysis. The Cogent NGS Discovery Software (CogentDS), an interactive data visualization tool, uses output files from CogentAP for downstream cluster analysis.