Scaling of nucleic acid assays on microelectrophoresis array devices: High‐dynamic range multi‐gene readout from less than ten transcripts

Abstract

In this paper we describe progress in using the prodigious data-collecting ability of multilane microelectrophoresis instruments to bear on problems in scaled nucleic acid assays. We emphasize compound stacking and Solid-Support loading as means to concentrate <100 pg samples for direct injection. Reaction Mapping is applied to readout of quantitative polymerase chain reaction (qPCR) gene-expression and as a way to practically overcome difficulty in interpreting amplification curves of multiplexed qPCR at 20-50 gene/well complexity. We demonstrate multiplexed readout of gene expression over an abundancy range of 9Log2 units starting with reverse-transcribed samples as small as 5 molecules in each sample.