Next-generation sequencing (NGS) library preparation faces three persistent challenges: 1) sample qualification uncertainties, 2) inefficient library purification/normalization processes, and 3) sequencing artifacts from PCR duplicates/adapter dimers. Current normalization methods relying on individual quantification (Qubit, Tapestation) and liquid handling robotics incur substantial costs, while failing to address sample-specific amplification needs. This proves particularly problematic for low-input FFPE samples where degradation and crosslinking create variable template quality undetectable by conventional quantification methods.

We present a solution powered by n6 iconPCR technology that enables:

• Real-time amplification monitoring through fluorescent tracer incorporation
• AutoNorm during PCR via cycle threshold adjustment
• Direct pooling from amplification plates

In FFPE samples, this approach:

• Reduced dropout rates
• Decreased PCR duplicates
• Eliminated pre-pooling quantification steps

This workflow enables reliable processing of challenging samples while reducing per-library costs through reagent and time savings. The technology's dynamic adaptation to sample quality parameters demonstrates particular value for degraded clinical specimens requiring maximal data recovery.