T7-based linear amplification of low concentration mRNA samples using beads and microfluidics

We have demonstrated in vitro transcription (IVT) of cDNA sequences from purified Jurkat T-cell mRNA immobilization on microfluidic packed beads down to single-cell quantities. The microfluidic amplified aRNA was nearly identical in length and quantity compared with benchtop reactions using the same starting sample quantities. Microarrays were used to characterize the number and population of genes in each sample, allowing comparison of the microfluidic and benchtop processes. Keywords: gene expression transcripts detected In this study we examined the IVT reaction in a microfluidic system apart from the other steps in the linear amplification process. Stock concentrations of cDNA were generated and known concentrations (down to single-cell equivalents) were packed onto the microfluidic or utilized as a benchtop process. IVT reactions were performed and efficiencies compared using microarray platforms.