mRNA isolation in a microfluidic device for eventual integration of cDNA library construction

摘要

mRNA isolation for the purpose of cDNA library construction was performed in a microfluidic chip device uing paramagnetic oligo-dT beads. The simple Y-intersection flow design mixes beads and sample on-chip, and uses magnetic trapping to capture, then release the beads. The capillary gel electrophoresis (CGE)detection of the total unamplified mRNA isolated on-chip, and of a reverse transcription-polymerase chain reaction (RT-PCR)amplified rare gene indicted that mRNA could be captured by oligo-dT beads on-chip, had very good integrity and was suitable for constructing a cDNA library. The limit of detection for the rare bicoid gene of Drosophila melanogaster corresponded to the capture of appoximately 2.8ng of total mRNA from 0.85 #mu#g of total RNA (TRNA)within the microchip. As much as 34 ng of total mRNA was estimated to be captured from 10 #mu#g of TRNA.