The SMART-Seq HT Kit is designed to generate high-quality, full-length cDNA directly from 1–100 cells or 10 pg–1 ng of total RNA in a high-throughput manner. The kit is evolved from the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (SMART-Seq v4) and features a simplified protocol, which combines the reverse transcription and PCR amplification into a convenient single step without compromising on performance. The user simply needs to set up the one-step RT-PCR and walk away until the double stranded cDNA is amplified and ready for purification.
The cDNA synthesis protocol can be completed in four hours, and the entire library construction protocol can be completed within two working days. Indeed, the SMART-Seq HT Kit has the same sensitivity and reproducibility that characterize the SMART-Seq v4 kit.
The core SMART technology in this kit is an immensely powerful tool for studying the full transcriptome, including splice junctions and alternative splicing events in cells, and provides full-length gene coverage. Oligo(dT) priming is used to specifically amplify mRNA from either high-quality total RNA (RIN >8) or intact cells. LNA technology improves the efficiency of template switching, which increases the number of genes identified to higher levels than any other SMARTer Ultra Low kit for mRNA-seq.