The Impact of Blood Sample Processing on Ribonucleic Acid (RNA) Sequencing

Area of Science:

• Molecular Biology
• Genomics
• Bioinformatics

Background:

• Gene quantification and expression analysis are vital in biological research.
• Sample selection and processing can introduce significant variables, potentially leading to ambiguous results.
• Understanding these impacts is crucial for accurate high-throughput sequencing data.

Purpose of the Study:

• To investigate the impact of sample storage time on RNA sequencing.
• To elucidate the mechanism by which sample processing affects gene expression analysis.
• To evaluate changes in RNA integrity and gene expression patterns under different storage conditions.

Main Methods:

• RNA was extracted from peripheral blood mononuclear cells (PBBMcs) and blood samples.
• RNA integrity was assessed after storage at room temperature for 0, 4, and 8 hours.
• High-throughput sequencing was performed for comprehensive gene expression analysis.

Main Results:

• Increased storage time led to greater differences in gene expression.
• The total number of detected genes remained largely unchanged with extended storage.
• Five novel protein-coding genes exhibited gradient expression patterns correlating with storage duration.

Conclusions:

• Sample storage time is a critical factor influencing gene expression variability in RNA sequencing.
• The study identified specific genes sensitive to storage conditions, offering new insights.
• This research underscores the importance of standardized sample processing protocols for reliable genomic studies.