New RNA-seq, metabolomics protocol offers more efficient extraction that maintains data integrity

The protocol employs a streamlined extraction from a single sample, which reduces variation, improves efficiency, preserves data fidelity and maximizes use of precious biospecimens.

“Our new technique enables researchers to study metabolic phenotypes in a unique way while getting the most information we can out of single samples,” said Ryan Sheldon, Ph.D., director of VAI’s Mass Spectrometry Core. “The most important takeaway is there is no loss of information by using our new, more efficient protocol.”

Until now, scientists have had to use two sample extractions — one for RNA-seq and one for metabolomics. This approach requires multiple samples or a single sample divided into subsamples, which increases variation and has an unknown impact on data interpretation.

The current workflow also can make multi-omics approaches challenging, especially in rare sample types; the extraction process destroys the sample, precluding additional analysis.

To validate the protocol, the research team compared results from the standard extraction to results from the new approach. The findings clearly showed the new protocol preserved data quality while saving time and resources.

The new protocol is published in the journal RNA Biology. It was developed by scientists in VAI’s Core Technologies and Services and Department of Metabolism and Nutritional Programming. Core Technologies and Services provides state-of-the-art technologies and expertise to Institute scientists and collaborators.

“Our new strategy offers an important proof-of-concept for future efforts to incorporate additional -omics approaches, with the goal of creating a singular extraction pipeline for RNA-seq, metabolomics, proteomics, transcriptomics, and others,” Sheldon said. “This work would not have been possible without the exceptional Core Technologies and Services team here at VAI and the Institute’s commitment to collaboration and rigorous science.”

Authors include Zachary B. Madaj, M.S., Michael S. Dahabieh, Ph.D., Vijayvardhan Kamalumpundi, Brejnev Muhire, Ph.D., Dean J. Pettinga, Rebecca A. Siwicki, M.S., Abigail E. Ellis, Christine Isaguirre, Martha L. Escobar Galvis, Ph.D., Lisa DeCamp, Russell G. Jones, Ph.D., Scott A. Givan, Ph.D., Marie Adams, M.S., of VAI. The Institute’s Mass Spectrometry Core, Pathobiology and Biorepository Core, Bioinformatics and Biostatistics Core and Genomics Core played key roles in protocol development.

Research reported in this publication was supported by Van Andel Institute.