High Quality Whole Transcriptome RNA-Sequencing Techniques

This technical report discusses RNA sequencing (RNA-seq) methodologies applied to challenging clinical samples, specifically tumor biopsies and whole blood. It outlines the difficulties encountered in RNA-seq, such as limited input material and high levels of ribosomal RNA, which can lead to compromised data quality. The report evaluates three RNA-seq workflows developed by WatchMaker Genomics, focusing on the effectiveness of the Polaris Depletion step in generating high-quality libraries. Results indicate that workflows incorporating Polaris Depletion yield low PCR duplication rates and higher gene detection, particularly for long non-coding RNAs. The performance of these workflows was consistent across formalin-fixed, paraffin-embedded (FFPE) and whole blood samples. The findings suggest that high-quality transcriptome data can be achieved without the need for exome capture, simplifying the workflow and reducing costs while maintaining data integrity. This study provides valuable insights into efficient RNA-seq approaches for clinical applications.