Takara SMART-Seq v4 full-length transcriptome analysis with ultimate sensitivity for ultra-low RNA inputs

NOTE: For RNA sequencing with ultra-low input, we recommend the reconfigured versions of the kits below, SMART-Seq mRNA LP (Cat. # 634768, 634769 & 634771) and SMART-Seq mRNA (Cat. # 634772, 634773 & 634770).

SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Cat. # 634888, 634889, 634890, 634891, 634892, 634893 & 634894) and SMART-Seq v4 PLUS Kit (Cat. # R400752 & R40753) will be phased out soon. Contact your local sales representative for more information.

NOTE: For RNA sequencing with ultra-low input, we recommend the reconfigured versions of the kits below, SMART-Seq mRNA LP (Cat. # 634768, 634769 & 634771) and SMART-Seq mRNA (Cat. # 634772, 634773 & 634770).

SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Cat. # 634888, 634889, 634890, 634891, 634892, 634893 & 634894) and SMART-Seq v4 PLUS Kit (Cat. # R400752 & R40753) will be phased out soon. Contact your local sales representative for more information.

The SMART-Seq v4 Kit (SSv4) and the SMART-Seq v4 PLUS Kit (SSv4 PLUS) are the fourth generation of our SMARTer ultra-low solutions and our most sensitive kits for ultra-low inputs. These kits use oligo(dT) priming to generate high-quality, full-length cDNA directly from multiple intact cells (up to 1,000 cells) or 10 pg–10 ng of total RNA. In addition to the cDNA synthesis kit, the SSv4 PLUS kit also includes a library preparation kit and a single-use unique dual index (UDI) plate to generate Illumina-compatible sequencing libraries, making it a complete end-to-end solution.

The SMART (Switching Mechanism at 5` End of RNA Template) technology powering the cDNA synthesis technology employed by the SMART-Seq v4 kit provides full-length transcript information, enabling analysis of transcript isoforms, gene fusions, point mutations, etc. Additionally, SSv4 takes this technology to the next level by improving upon the Smartseq2 method and incorporating locked nucleic acid (LNA) technology in the SMART-Seq v4 Oligo for more efficient template switching. This allows for the identification of higher numbers of genes relative to other methods. The cDNA kits provide high reproducibility, even gene-body coverage, and an accurate representation of GC-rich transcripts.

The SSv4 PLUS kit includes our patented library preparation kit that incorporates enzymatic fragmentation and stem-loop adapters to construct high-quality, Illumina-compatible libraries from the cDNA synthesized with the core kit. This two-step workflow takes place in a single tube and can be completed in about two hours with no intermediate purification steps necessary—minimizing sample mix-up, sample loss, and cross-contamination. The high library yield generated from this kit provides the flexibility to sequence on high-throughput sequencers while providing high reproducibility.

If you are interested in these benefits but are working with intact, high-quality single cells, we recommend our SMART-Seq Single Cell PLUS Kit.

If you prefer a random priming approach that will allow you to work with degraded samples and also includes library preparation and indexing reagents, we recommend our SMART-Seq Stranded Kit.

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Overview

• Single-tube workflows—complete cDNA synthesis and library preparation, in one tube each, for minimization of sample loss, sample mix-up, or other handling errors
• Unparalleled sensitivity—start with as little as 10 pg of total RNA (input range: up to 1,000 cells or 10 pg–10 ng of total RNA)
• High-quality RNA-seq data—identify the highest number of genes for total RNA; obtain full-length transcript information, a low percentage of rRNA reads, and a great representation of GC-rich transcripts
• Increase your sequencing power—include unique dual indexes to allow for the pooling of multiple samples and confident sequencing on the NovaSeq™ system
• High library yield —sequence on high-throughput sequencers with ease (such as the NovaSeq system) or confidently save extra library for future runs

Interested in more data and FAQs about this product? Visit the NGS Learning Center.

Figure 1. SMART-Seq v4 PLUS generates more libraries and identifies more genes than Nextera® XT. Panel A. SSv4 chemistry was used to produce cDNA, in triplicate, from 10 pg of Mouse Brain Total RNA. Illumina-compatible libraries were then generated using the SMART-Seq Library Prep Kit (not sold separately) portion of the SSv4 PLUS kit or Nextera XT and sequenced on a NextSeq® 500. The reads were normalized to 4M paired-end reads and analyzed as described in the techical note. Panel B. The library yield obtained with the SSv4 PLUS kit is higher than the yield obtained when using Nextera XT. Panel C. The number of genes identified was also higher for the SSv4 PLUS kit compared to Nextera XT.

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Applications

• cDNA synthesis from multiple cells or total RNA for transcriptome sequencing
• cDNA outputs from the SSv4 kit can be used with either the SMART-Seq Library Prep kit (not sold separately; sold as part of the SSv4 PLUS kit) for Illumina sequencing or with the Ion Xpress Plus Fragment Library Kit for Ion Torrent sequencing

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Product Components List to determine kit components. Certificates of Analysis and Product Components Lists are located under the Documents tab.