SMART-Seq mRNA Single Cell LP and SMART-Seq mRNA Single Cell
The SMART-Seq mRNA Single Cell Kit (SSmRNA Single Cell) and the SMART-Seq mRNA Single Cell LP Kit (SSmRNA Single Cell LP) are powered by robust chemistry that provides unparalleled sensitivity and reproducibility for single-cell and nuclei applications. These kits use oligo(dT) priming to generate high-quality, full-length cDNA directly from single cells known to have low RNA content (e.g., PBMCs, T cells, B cells, etc.). Using the SSmRNA Single Cell LP kit with Unique Dual Index kits (sold separately), up to 384 multiplexed Illumina-ready sequencing libraries can be generated. The kit offers an end-to-end solution from sample to cDNA synthesis, library preparation, and data analysis with our free Cogent NGS bioinformatics pipeline.
The SMART (Switching Mechanism at 5′ End of RNA Template) technology powering SSmRNA Single Cell provides full-length transcript information, enabling analysis of transcript isoforms, gene fusions, point mutations, etc. The cDNA kit provides high reproducibility, even gene-body coverage, and an accurate representation of GC-rich transcripts. We have further modified this powerful core technology to create a new chemistry with higher sensitivity designed specifically for single-cell applications. These enhancements result in more useful reads and a higher gene detection, allowing for a better understanding of precious samples.
The SSmRNA Single Cell LP 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 cells (<1,000) or ultra-low total RNA inputs, we recommend our SMART-Seq mRNA LP Kit or our SMART-Seq mRNA LP (with UMIs) Kit.
If you prefer a random priming approach that will allow you to work with degraded samples and includes library preparation and indexing reagents, we recommend our SMART-Seq Stranded Kit.
Overview
- Easy, plate-based workflow—direct input of single cells isolated by FACS or other methods
- 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 and reproducibility—get the best performance for single-cell RNA-seq (especially for cells with very low RNA content and for nuclei) with the highest gene detection
- Robust, full-length chemistry—outperform all existing single-cell full-length methods, including Smart-seq2, with this validated protocol
- Increased 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
- Highly scalable workflow with flexible, miniaturized volumes—automate and use for high-throughput applications
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Data is shown for the SMART-Seq Single Cell Kit (SSsc) and SMART-Seq Single Cell PLUS (SSsc PLUS), which use the same chemistry as SMART-Seq mRNA Single Cell and SMART-Seq mRNA Single Cell LP.
SMART-Seq Single Cell Kit provides higher sensitivity than Smart-seq2
Validated kit backed by expert technical support
Figure 1. The SMART-Seq Single Cell Kit outperforms Smart-seq2. Single cells from the lymphoblastoid cell line GM12878 were processed with the SMART-Seq Single Cell Kit (SSsc; 18 cells) or the Smart-seq2 method (Smart-seq2; 20 cells) using 19 cycles of PCR. RNA-seq libraries were generated and sequences analyzed (after normalizing all samples to 1.75 million paired-end reads). Panel A. The read distribution was different between the two chemistries, with a drastically higher number of reads mapping to the mitochondrial genome (and therefore, fewer reads available for gene identification) with Smart-seq2 chemistry. Panel B. More genes were detected in the cells processed with SSsc. For additional data on reproducibility and sensitivity, view our technical note.
Even higher sensitivity for single-cell applications
We have further modified our core technology to create a chemistry with higher sensitivity designed specifically for single-cell applications. We demonstrated that the SSsc kit generates data with even better sensitivity and reproducibility than a kit using our core technology (SSv4).
Figure 2. The SMART-Seq Single Cell Kit's improved chemistry is highly suitable for single-cell applications. Panel A. 12 single cells from lymphoblastoid cell line GM22601 were processed with SSv4 or SSsc using 19 cycles of PCR. All samples were normalized to 1.25 million paired-end reads. The cDNA yield generated with SSsc was drastically higher than that generated with SSv4. Panel B. About 50 single PBMCs from one donor were processed with SSv4 or SSsc. About 60% more genes were detected in the cells processed with SSsc, regardless of the number of reads used for the analysis.
SSsc PLUS provides a complete workflow from cDNA through library preparation
The SMART-Seq Single Cell PLUS Kit is designed to synthesize high-quality, full-length cDNA directly from single cells, and then generate Illumina-ready sequencing libraries.
Figure 3. SSsc PLUS generates more libraries and identifies more genes than Nextera® XT. Panel A. The SMART-Seq Single Cell Kit was used to produce cDNA, in triplicate, from 10 pg of Mouse Brain Total RNA. Illumina-compatible libraries were then generated using either the SMART-Seq Library Prep Kit portion of the SSsc PLUS kit or Nextera XT. Libraries were sequenced on a NextSeq® 500. The reads were normalized to 4M paired-end reads and analyzed as described in the technical note. Panel B. Library yields obtained with the SSsc PLUS kit are higher than yields produced with Nextera XT. Panel C. The SSsc PLUS kit also identified more genes compared to Nextera XT.
More Information
Applications
- cDNA synthesis from single cells for full-length transcriptome sequencing
- cDNA outputs from SMART-Seq mRNA Single Cell can be used with SMART-Seq Library Prep (not sold separately; sold as part of SMART-Seq mRNA Single Cell LP) for Illumina 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.
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Uncover the full tale of your cells
Wondering how well you’ve truly studied your single-cell sample? Worry no more! The SMART-Seq Single Cell Kit's full-length chemistry lets you capture entire transcripts—giving you the power to identify rare genes and detect SNPs, fusions, and more—filling in the gaps that 3’ methods, like 10x, can miss.
Complete end-to-end RNA-seq workflows
Our SMART-Seq PLUS kits have it all. With just one kit, go from sample to sequencing-ready library.
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