Technological Validation of Single Cell Expression Analysis

Toulouse « Genome and Transcriptome » core facility (France) bought and validated Fluidigm C1 Single-Cell auto prep system. This technology separates and prepares up to 96 individual cells for transcriptomic analysis. The workflow is : mRNA extraction from individual cells, reverse transcription and pre-amplification (Specific Target Amplification). The final preamplified cDNA product is thereafter harvested to collection wells for transfer to the BioMark HD System for quantitative PCR analysis. We performed a technical validation on : reproducibility, absence of contamination, detection of the viability of the injected cells, percentage of missing data, correlation between single cell expression median and cellular pool  expression (used as positive control) on 96 genes. Results obtained during this validation step are promising.