High Quality RNA Sequencing Method Evaluation

This technical report evaluates the performance of the Watchmaker Genomics RNA Library Prep with Polaris Depletion workflow for RNA sequencing of challenging clinical samples. It addresses the difficulties associated with RNA sequencing, such as limited sample material, RNA fragmentation, and high levels of ribosomal RNA and globin RNA, which can lead to increased PCR duplication rates and compromised data quality. The report compares this new workflow to a standard RNA-seq and exome capture method across various sample types, including universal human reference RNA, whole blood, and tumor FFPE samples. Key findings indicate that the Polaris Depletion workflow significantly reduces PCR duplication rates and improves gene detection and expression quantification. The results demonstrate that high-quality RNA-seq libraries can be generated efficiently, enhancing sensitivity and workflow efficiency without the need for costly exome capture. The report includes detailed performance metrics and comparisons across different RNA qualities and input amounts.