Differential expression analysis on the ICELL8 cx Single-Cell System
By leveraging Takara Bio's expertise in single-cell RNA-seq library construction and the cell isolation and selection capabilities of the ICELL8 platform, the ICELL8 cx 3' DE workflow enables high-throughput preparation of Illumina libraries for differential expression analysis of single cells from up to eight different samples in parallel.
By leveraging Takara Bio's expertise in single-cell RNA-seq library construction and the cell isolation and selection capabilities of the ICELL8 platform, the ICELL8 cx 3' DE workflow enables high-throughput preparation of Illumina libraries for differential expression analysis of single cells from up to eight different samples in parallel.
The ICELL8 cx 3' DE workflow for the ICELL8 cx Single-Cell System uses the ICELL8 3' DE Reagent Kit (Cat. # 640167); the ICELL8 cx 3' DE Chip (Cat. # 640199), a 5,184-nanowell SmartChip that contains preprinted oligo(dT) primers, and reagents for cDNA synthesis and Illumina library prep; the ICELL8 cx Loading Kit (Cat. # 640197); and the ICELL8 Collection Kit – L (Cat. # 640212)—all purchased separately.
Following staining and dilution, sample cells are dispensed into chip nanowells using the MultiSample NanoDispenser (MSND) part of the cx system. Wells containing single, viable cells are selected using the ICELL8 cx Imager and the ICELL8 cx CellSelect Software provided with the ICELL8 cx system. Following the selection of cells for processing and analysis, RT-PCR reagents are dispensed into the corresponding wells, and cDNA synthesis and amplification are performed in-chip in a single run. Following RT-PCR, cDNA products are pooled and purified, and sequencing libraries are prepared using a transposase-based method that enriches for cDNA derived from mRNA 3' ends, followed by a limited-cycle PCR amplification step that incorporates Illumina index sequences.
Sequencing data from the resulting libraries can be traced back to individual cells using the nanowell-specific barcode sequences and can be used to perform differential gene expression analysis by mRNA 3' end-counting. By yielding high-quality, 3'-focused sequencing data from hundreds of cells in parallel, the kit provides an efficient, cost-effective solution for high-throughput differential expression analysis of single cells.
Overview
- High-throughput single-cell analysis—generate sequencing data from hundreds of single cells in parallel
- Sample flexibility—analyze cells from up to eight different samples in a single experiment
- mRNA 3' sequence enrichment—reduce the number of reads required for differential gene expression analysis by enriching for sequences derived from mRNA 3' ends
- Reduced sample handling—minimize variability with automated cell isolation and selection, and sample pooling prior to library construction
- Illumina sequencing libraries—generate libraries suitable for sequencing on all Illumina platforms
More Information
Components
The following items are required to complete the workflow:
ICELL8 cx Single-Cell System (Cat. # 640188 or # 640189) | ||
---|---|---|
Cat. # | Product | 1 item required per: |
640167 | ICELL8 3' DE Reagent Kit | 1 chip |
640199 | ICELL8 cx 3' DE Chip | 1 run |
640212 | ICELL8 Collection Kit – L | 1 chip |
640197 | ICELL8 cx Loading Kit | 1 chip |
640192 | ICELL8 384-Well Source Plate and Seal (5/pack) | 1 chip |
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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