SureCell WTA 3′ Library Prep Kit for the ddSEQ System
A robust, scalable, and user-friendly workflow for transcriptome profiling of hundreds to tens of thousands of single cells in a given experiment.Read More...
The Illumina Bio-Rad® SureCell™ Whole Transcriptome Analysis (WTA) 3’ Kit provides a validated process for single-cell RNA sequencing studies.
Single cells are individually partitioned into subnanoliter droplets on a disposable cartridge on the one-touch ddSEQ™ Single-Cell Isolator. The cartridge can accommodate multiple samples, and multiple cartridges can be processed in parallel to isolate thousands of cells in a matter of minutes.
Cell lysis and cell barcoding occur inside individual droplets, and single-cell-barcoded RNA-Seq libraries are subsequently prepared using Nextera technology.
Conduct data analysis via BaseSpace Sequence Hub, the Illumina cloud-based genomics computing environment.
View sample data (requires a BaseSpace login)
This kit has been validated for human and mouse samples. The BaseSpace Sequence Hub SureCell RNA Single-Cell App supports analysis of human, mouse, rat, zebrafish, C. elegans, and Drosophila samples.
| Instrument | Recommended Number of Samples | Read Length |
|---|---|---|
| NextSeq 550 System | 4 to 6 samples per run (based on a minimum of 100,000 reads per cell) | read 1: 68 bp, read 2: 75 bp |
| HiSeq 2500 System | Rapid run: 4-5 samples per lane; high output v4: 4 samples per lane (based on a minimum of 100,000 reads per cell) | read 1: 68 bp, read 2: 75 bp |
| SureCell WTA 3′ Library Prep Kit for the ddSEQ System | TruSeq RNA Library Prep Kit v2 | Illumina Stranded mRNA Prep | Illumina RNA Prep with Enrichment |
|---|
Mouse cell lines (A20, NIH3T3) and human cell lines (HEK, BJ) were processed using the SureCell WTA 3′Library Prep Kit. Consistently high numbers of detected genes demonstrates that recovery of transcripts is not limited by cell size.
A. BaseSpace-generated plots of the number of unique molecular identifiers (UMI), i.e. transcripts, assigned to the mouse/mm10 (red) and human/hg19 (blue) genomes for each cell barcode. Unique transcripts mapping to both mouse and human (purple) represent cell doublets. B. Cumulated fraction of unique transcripts assigned to cell barcodes (linear scale). The inflection point (red line) determines the number of barcoded cells detected in the sequencing run.
A. PCA clustering of 1384 single cells sorted from a 1:1 ratio mixture of HEK 293 and NIH 3T3 cells, sequenced and analyzed with the BaseSpace SureCell Single-Cell RNA App. B. Cell populations cluster based on expression of the human (hg19) RPL13 gene.
A. Analysis in the BaseSpace SureCell Single-Cell RNA App using the t-SNE algorithm of a mixture of NIH 3T3 and HEK 293 cells identifies a distinct subpopulation of cells. B. Cells color coded by gene expression of hg19 RPL 13 confirms the identity of the subpopulation as human.
Analysis of cell cycle state using the BaseSpace SureCell Single-Cell RNA App is based on unique transcript counts of genes associated with each phase of the cell cycle, normalized by the total count for each cell. Expression is centered by the median and scaled by the median absolute deviation for each cell cycle.
Single-Cell RNA Data Analysis Workflow
Technical Note | PDF< 1 MB
Illumina | Bio-Rad Single-Cell RNA Sequencing of Peripheral Blood Mononuclear Cells
Application Note | PDF1 MB
Illumina Bio-Rad Single-Cell RNA Sequencing Solution
Data Sheet | PDF< 1 MB
SureCell WTA 3' Library Prep Kit Documentation
Custom Protocol Selector
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