CryoGrid-PIXUL-RNA: high throughput RNA isolation platform for tissue transcript analysis

Area of Science:

• Biotechnology
• Genomics
• Molecular Biology

Background:

• High-throughput multiomics requires efficient tissue analysis workflows.
• Current biospecimen storage and sampling methods are inefficient, creating a bottleneck.
• The PIXUL sonicator facilitates analyte extraction from cells and tissue fragments.

Purpose of the Study:

• To develop a user-friendly system for cataloging, cryostoring, and sampling tissue fragments.
• To create a streamlined, high-throughput RNA isolation protocol for tissue analysis.
• To integrate a novel RNA isolation method with existing tissue processing technologies.

Main Methods:

• Development of the CryoGrid system for tissue cryostorage and sampling.
• Creation of a microplate-based, TRIzol-free, and column-free RNA isolation protocol using proteinase K (PK) buffer.
• Integration of the CryoGrid system with the PIXUL sonicator and various RNA isolation methods (PK buffer, TRIzol, PureLink columns).

Main Results:

• The CryoGrid-PIXUL system, combined with PK buffer, TRIzol, or PureLink columns, yielded comparable RNA transcript profiles.
• Transcript profiles from frozen mouse organs (brain, heart, kidney, liver) in a sepsis model were analyzed.
• Demonstrated successful gene-specific qPCR and genome-wide transcript analyses.

Conclusions:

• The CryoGrid-PIXUL system with the 96-well microplate PK buffer method provides an inexpensive, user-friendly, high-throughput workflow.
• This integrated system enables efficient study of transcriptional responses in tissues.
• Facilitates research in various conditions including health, disease, and therapeutic interventions.