Dr. Esser’s group at the University of Florida previously showed that cellular circadian clocks direct a daily transcriptional program that supports homeostasis and resilience. To further learn the age-associated changes in circadian functions, they profiled the circadian transcriptome in different tissues including skeletal muscle, the heart, and others. This study has more than 440 samples.
To speed up the library construction and potentially save reagents cost, we manually tested ½x, ¼x, 1/5x and 1/10x reduced reaction volumes for RNA-seq library construction using NEB Ultra Directional RNA-seq kit. The results showed that 1/5x gave us reliable results. Thus, by leveraging the mosquito® and dragonfly® discovery automated liquid handling systems for mRNA-seq library construction with 1/5x reduced reaction, reagent costs were reduced five-fold via miniaturization whilst maximizing the reliability of an automated workflow.
Key learning objectives
- Explore the first-hand experience of automation-enabled miniaturization of mRNA-seq
- Learn about the first-hand experience of the utility of mosquito and dragonfly discovery for NGS in a shared core facility
- Understand the fundamental mechanism of molecular clocks and their impact on physiology, using skeletal muscle as a tissue focus
- Discover the application of sequencing tools to investigate the influence of circadian rhythms, particularly with aging
