Preservation of scRNA-Seq Libraries Using Existing Inactivation Protocols.

Area of Science:

• Molecular Biology
• Virology
• Genomics

Background:

• Single-cell RNA sequencing (scRNA-seq) is a powerful tool for understanding cellular responses in disease and infection.
• High-containment laboratories (Biosafety Level 3 [BSL-3] and BSL-4) are essential for studying dangerous pathogens but pose challenges for scRNA-seq.
• Standard inactivation protocols for sample processing in containment may affect the integrity of sensitive nucleic acid libraries.

Purpose of the Study:

• To evaluate the feasibility of removing scRNA-seq libraries from high-containment settings using established nucleic acid inactivation methods.
• To determine if common inactivation buffers (Trizol, RLT, AVL) compromise the quality of scRNA-seq libraries.

Main Methods:

• Single-cell RNA sequencing libraries were processed using standard inactivation protocols (Trizol, RLT buffer, AVL buffer).
• The quality and integrity of the inactivated libraries were assessed using established metrics.
• Comparative analysis was performed between inactivated and non-inactivated samples.

Main Results:

• Existing inactivation protocols for nucleic acid extraction were demonstrated to be effective for scRNA-seq libraries.
• Inactivation using Trizol, RLT buffer, or AVL buffer did not significantly affect sample quality or library integrity.
• The tested inactivation methods are compatible with the requirements for downstream scRNA-seq analysis.

Conclusions:

• It is feasible to remove scRNA-seq libraries from high-containment laboratories using existing nucleic acid inactivation protocols.
• These findings provide a validated workflow for safely handling and processing scRNA-seq samples from BSL-3/BSL-4 environments.
• The study supports the broader application of scRNA-seq for studying high-containment pathogens without compromising data quality.