RNA sequencing depth guidelines for the study of alternative splicing.

Deep, paired-end RNA-seq analysis in human buffy coat cells and multiple tissues shows typical published sequencing depths are not sufficient to comprehensively capture alternative splicing.

• Why it matters: Budget limits force a trade-off between sample number and sensitivity, especially for lowly expressed genes, yet splicing depth guidance has been limited to human adipose tissue.
• What they did: The study analyzed publicly available and newly generated deep-sequenced paired-end RNA-seq data (150 to >500 million reads; 50–150 bp) across human buffy coat cells and tissues including gluteal subcutaneous fat, heart, and hypothalamus.
• The result: The findings motivate deeper sequencing or long-read technologies and lead to practical sequencing-depth guidelines plus a website to select an appropriate depth for alternative splicing studies.

AI summary of the abstract · may contain errors · not medical advice · read the paper.