SEQURNA enhances FLASH-seq gene detection while eliminating DTT dependence
Effective RNase inhibition is critical for single-cell RNA-sequencing, yet commercial recombinant RNase inhibitors (RRIs) require reducing agents for stability and impose substantial costs. Here, we systematically benchmark SEQURNA, a synthetic thermostable RNase inhibitor, against commercial alternatives using FLASH-seq in human retinal organoids and peripheral blood mononuclear cells (PBMCs). SEQURNA at 0.5-1 U/ul achieved 14-25% higher gene detection than Takara RRI, with the greatest improvement in low-RNA PBMCs. Surprisingly, DTT supplementation at standard concentrations (5-10 mM) significantly impaired gene detection across all SEQURNA concentrations without improving RNA quality metrics, challenging established reverse transcription protocols. SEQURNA preserved biological heterogeneity, maintained sample stability during one-month storage at -80 degrees C, and reduced reagent costs by over 75%. We recommend SEQURNA at 0.75 U/ul without DTT as an optimized formulation that simultaneously enhances data quality and cost-effectiveness for full-length single-cell RNA sequencing.