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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 3" DE and ICELL8 3" DE for UMI workflows enable 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 ICELL8 systems, the ICELL8 3" DE and ICELL8 3" DE for UMI workflows enable high-throughput preparation of Illumina libraries for differential expression analysis of single cells from up to eight different samples in parallel.
The ICELL8 Chip and Reagent 3" DE Kit (Cat. # 640164, 640165, 640166) is compatible with the original ICELL8 Single-Cell System. Included in the kit are an ICELL8 3" DE Chip (Cat. # 640143), a 5,184-nanowell SmartChip that contains preprinted oligo(dT) primers, and reagents for cDNA synthesis and Illumina library prep.
Following staining and dilution, sample cells are dispensed into chip nanowells using the ICELL8 MultiSample NanoDispenser (MSND). Wells containing single, viable cells are selectedusing the ICELL8 Imaging System and the CellSelect Software provided with the ICELL8 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, these kits provide an efficient, cost-effective solution for high-throughput differential expression analysis of single cells.
The ICELL8 3" DE for UMI Reagent Kit is used with the ICELL8 3" DE Chip as part of the ICELL8 3" DE with UMI workflow, which enables Illumina sequencing and differential gene expression analysis-via transcript 3" end counting-of single cells isolated on the ICELL8 system.
