Single-cell full-length transcriptome analysis on the ICELL8 system powered by SMART-Seq technology
Full-length sequence information allows many types of analysis—such as differential gene expression, gene fusion, and alternative splicing—with high sensitivity. Our high-throughput solution leverages SMART-Seq chemistry for sensitive, full-length transcriptome profiling of 1,000–2,000 cells isolated on the ICELL8 Single-Cell System.
Please refer to the table in the "Components" section below for ordering information.
Full-length sequence information allows many types of analysis—such as differential gene expression, gene fusion, and alternative splicing—with high sensitivity. Our high-throughput solution leverages SMART-Seq chemistry for sensitive, full-length transcriptome profiling of 1,000–2,000 cells isolated on the ICELL8 Single-Cell System. The workflow incorporates Illumina-specific adapter sequences during cDNA amplification on an ICELL8 chip for the generation of indexed libraries that are ready for sequencing on Illumina platforms.
Please refer to the table in the "Components" section below for ordering information.
Single cells isolated with the ICELL8 Single-Cell System (Cat. # 640000) are dispensed into a 5,184-nanowell, blank ICELL8 250v Chip (Cat. # 640183). Following identification of single-cell-bearing wells using CellSelect Software, SMART RT-PCR reagents are then dispensed to the wells of samples chosen for further processing. The chip is then transferred to the ICELL8 Chip Cycler, which is programmed to perform one-step RT-PCR for first-strand synthesis and amplification of cDNA in a single run. The full-length cDNA is tagmented by TDE1 (Illumina Tagment DNA Enzyme), and then the tagmented cDNA is amplified using forward and reverse indexing primers (Cat. # 640205 and/or 640218). The resulting library is extracted, purified, and amplified to yield the final sequencing-ready library.
Overview
- High-throughput single-cell analysis—generate sequencing data from >1,000 single cells in parallel
- Multiplex for higher throughput—use SMART-Seq ICELL8 indexing primer sets A and B on two chips, and then pool the samples to allow sequencing of >2,000 cells simultaneously
- Sample flexibility—analyze cells from up to eight different samples in a single experiment
- Reduced sample handling—minimize variability with automated cell isolation and selection, and sample pooling prior to sequencing-ready library construction
- Illumina sequencing libraries—generate libraries suitable for sequencing on all Illumina platforms
More Information
Applications
- High-throughput, full-length single cell RNA-seq library preparation for Illumina platforms
Components
The following Takara Bio items are required to complete the workflow. Please refer to the SMART-Seq ICELL8 Application Kit User Manual for a complete list of materials required.
The following kits contain the necessary chip(s), reagents, and consumables required to run one, three, or five experiments (respectively) of single-cell, full-length transcriptome analysis.
| ICELL8 Single-Cell System (Cat. # 640000) | ||
|---|---|---|
| Cat. # | Product | Number of experiments per kit |
| 640219 | SMART-Seq ICELL8 Application Kit – 1 Chip1 | 1 |
| 640220 | SMART-Seq ICELL8 Application Kit – 3 Chip1 | 3 |
| 640221 | SMART-Seq ICELL8 Application Kit – 5 Chip1 | 5 |
| SMART-Seq ICELL8 application kit components | ||||
|---|---|---|---|---|
| Cat. # | Component2 | 640219 (1 Chip) |
640220 (3 Chips) |
640221 (5 Chips) |
| 640183 | ICELL8 250v Chip | 1 | 3 | 5 |
| 640212 | ICELL8 Collection Kit – L | 1 | 3 | 5 |
| 640206 | ICELL8 Loading Kit – B | 1 | 3 | 5 |
| 640202 | SMART-Seq ICELL8 Reagent Kit | 1 | — | — |
| 640203 | SMART-Seq ICELL8 Reagent Kit | — | 1 | — |
| 640204 | SMART-Seq ICELL8 Reagent Kit | — | — | 1 |
| 640205 | SMART-Seq ICELL8 Indexing Primer Set – A | 1 | 2 | 3 |
| 640218 | SMART-Seq ICELL8 Indexing Primer Set – B | — | 1 | 2 |
Notes:
- The application kits contain Cat. # 640212 (ICELL8 Collection Kit – L), which must be used with the ICELL8 Extraction Fixture – L (Cat. # 640217, purchased separately). If needed, please either log in to your research or business account and use 640217 to quick order as a one-time purchase or contact Customer Service to order one.
- Components can also be purchased separately.
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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Improve detection of fusions, SNPs, and alternative splicing with the ICELL8 system
We adapted our trusted SMART-Seq chemistry on the ICELL8 system for improved full-length scRNA-seq, so you can:
- Fully automate the processing of >1,000 individual cells into sequencing-ready libraries
- Achieve uniform coverage from 5'→3'
- Improve detection of gene fusions, SNPs, and splice variants
Read the technical note to see how our method compares to a 3'DE- and droplet-based method.
Read more
Single-cell identification with CellSelect Software
See how the ICELL8 platform's Imaging System and CellSelect Software allow for high-fidelity identification of individual cells.
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