Miniaturization and scale-down of reagent and reaction volumes for low-input RNA-Seq workflows enables researchers to work with low sample input and increased numbers of samples. As the reaction volumes are reduced, accurate delivery of reagent volumes emerges as a critical step to maintain data quality and technical reproducibility.
Researchers from Formulatrix and Clontech Laboratories present results from miniaturization of a low input RNA kit for generation of high-quality RNA-Seq samples using a compact desktop microfluidic dispensing technology. Reducing reagent volumes for cDNA synthesis using this kit provides results comparable to fullscale reaction volumes when using this microfluidic dispensing system. Average yield of amplified cDNA from the scaled-down reaction volume workflow is compatible with input samples of cells and total RNA. RNA-Seq data analysis metrics are comparable between the full-scale and miniaturized reaction conditions. The combination of high-yield cDNA synthesis chemistry and high-precision low-volume reagent dispensing provides a workflow well-suited for large-scale RNA-Seq studies.