Single-cell RNA sequencing (scRNA-seq) library preparation has long faced a critical bottleneck: choosing a fixed PCR cycle number before knowing how each sample will behave. When inputs vary, this forces researchers to split samples across multiple runs, quantify libraries between PCR steps, and risk over- or under-amplification — adding hands-on time, workflow complexity, and reagent waste while introducing artifacts that obscure the biology.
A Smarter Approach to PCR Amplification
n6’s icon96™ with AutoNorm™ technology replaces the fixed-cycle approach with real-time PCR control. Each reaction is monitored individually and automatically stopped at the optimal point in the amplification curve, eliminating the need for pre-quantification or batching by cell number.
Key Performance Highlights
Compared to a standard 10x Genomics workflow, icon96™ with AutoNorm™ delivers:
- Simultaneous processing of samples ranging from 500 to 10,000 cells, reducing thermocycler runs
- Fewer total PCR cycles per sample, minimizing overamplification
- Elimination of pre-index PCR quantification for faster turnaround
- 40–60% reduction in hands-on time
- Sequencing-ready libraries with no compromise in data quality
Consistent Data Quality Across Variable Inputs
n6’s application note demonstrates that single-cell gene expression data quality is maintained across all input ranges tested, with clustering results showing no meaningful difference between the icon96™ protocol and the standard workflow. The simplification is achieved without sacrificing scientific integrity.
This approach is particularly valuable for laboratories processing heterogeneous samples where cell numbers and RNA content vary between runs — a common reality in translational and clinical research settings.
Real-time PCR control allows each reaction to find its own optimal endpoint, removing the guesswork that has historically made scRNA-seq library prep labor-intensive and error-prone.
For research teams looking to increase throughput, reduce variability, and streamline their single-cell workflows, icon96™ with AutoNorm™ represents a meaningful step forward in library preparation technology.