- Measurement of expression levels of genes, alleles and spliced variants,
- Comparison of expression profiles between individual cells and populations,
- Mapping of transcription initiation sites,
- Characterization of alternate splicing patterns,
- Evaluation of post-transcriptional activity, and
- Discovery of new transcripts and gene fusions.
Fluidigm Corporation (NASDAQ:FLDM) today introduced the mRNA sequencing application on its C 1™ Single-Cell Auto Prep System. With this new C 1 system application, researchers have an easy, end-to-end workflow for detailed transcriptome analysis of 96 single cells to rapidly study differential expression profiles of diverse cell populations. The C 1 mRNA Seq Kits and integrated fluidic circuits (IFCs) enable the parallel processing of up to 96 single-cell cDNA libraries for quantitation of mRNA expression on Illumina sequencers. The protocol and reagent kits are expected to be commercially available in January 2013. Increasingly, researchers are analyzing the transcriptome at greater depth to uncover new mechanisms of cell development, metabolism and disease. mRNA sequencing has become a valuable tool to help researchers understand how cell sub-populations respond to signals and other environmental cues at critical stages of cell-fate determination or when they acquire aberrant phenotypes. Studying these gene expression patterns in single cells already has dramatically advanced cell biology. Unfortunately, most standard methods, such as microarray profiling or next-generation sequencing, are impractical for single-cell analysis because those technologies require large numbers of cells and are based on complex workflows that are too slow or generate highly variable results. The C 1 mRNA sequencing workflow was specifically optimized for high throughput single-cell analysis. All aspects of the workflow, including cell capture, staining for cell viability, lysis, reverse transcription and amplification are consolidated on C 1 system IFCs. The C 1 system workflow requires as few as 200 cells, utilizes as little as 10 picograms per cell, and requires no reagent mixing, fragmentation or purification. Researchers use the BioMark™ HD system to conduct routine quality control on the cDNA library in an effective and cost-efficient manner before running it on an Illumina sequencing system. This provides a reliable and standardized workflow for global gene expression analysis. “mRNA sequencing is a powerful tool to sequence and quantify all messenger RNA in an unbiased and hypothesis-free approach. However, very few laboratories have been able to conduct single-cell mRNA sequencing studies in a meaningful and reproducible way. By enabling this application on the Fluidigm C 1 system, we’ve simplified the workflow to only three hours of hands-on time and at one-eighth the cost of other library preparation methods,” said Candia Brown, Fluidigm’s Director of Product Marketing in the Single-Cell Genomic Business. “By automating the workflow and reducing the sample preparation cost, we are enabling virtually any laboratory to run larger scale, transcriptome-wide studies.” The C 1 Single-Cell mRNA sequencing application enables: