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.
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.
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.
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.