Multi-well plate lid for single-step pooling of 96 samples for high-throughput barcode-based sequencing.

Area of Science:

• Molecular Biology
• Genomics
• Biotechnology

Background:

• High-throughput transcriptomics, particularly RNA sequencing (RNA-seq), is crucial for biological and clinical research, driving biomarker and treatment development.
• Current RNA-seq workflows face limitations in high-throughput scalability due to cumbersome sample preparation steps.
• Bulk RNA barcoding and sequencing (BRB-seq) aims to improve throughput but requires optimized sample preparation.

Purpose of the Study:

• To develop and validate a novel pooling technology for streamlined sample preparation in BRB-seq workflows.
• To address the limitations of manual sample handling and reagent consumption in high-throughput RNA sequencing.
• To enable cost-efficient and large-scale sample processing for transcriptomic analysis.

Main Methods:

• Development of a smart labware pooling lid with integrated fluidic features for standard 96-well plates.
• Demonstration of rapid (under one minute) and high-efficiency (over 90%) liquid recovery in a single pooling step.
• Transition of the pooling lid from a prototype to large-scale injection molded manufacturing.

Main Results:

• The pooling lid successfully enabled simultaneous pooling of 96 samples from a multi-well plate into a single tube.
• The technology demonstrated efficient pooling of DNA barcodes for subsequent sequencing, validating its utility in BRB-seq.
• The system significantly reduces reagent consumption and manual labor compared to traditional methods.

Conclusions:

• The developed pooling lid is a valuable tool for enhancing the throughput of BRB-seq workflows.
• This innovation offers a cost-effective solution for large-scale sample preparation in transcriptomics.
• The technology has the potential to accelerate biomarker discovery and therapeutic development through efficient RNA sequencing.