The integrity of the sample material has a critical impact on the analysis results. Degraded RNA can lead to incomplete or inaccurate sequencing data, as fragmented RNA may not be efficiently captured or aligned. This affects gene-body coverage, resulting in uneven sequencing depth across the gene. Degraded RNA typically shows reduced coverage, particularly in the 3' and 5' regions of genes, leading to biased or incomplete gene expression profiles. High-quality, intact RNA is essential for consistent gene-body coverage and reliable, reproducible RNA sequencing results.
In particular, for poly-A enrichment, degraded RNA can introduce a strong bias towards the 3' end of the transcript. For these samples, we recommend rRNA depletion to improve data quality and reduce this bias.