icon96™

This application note describes how adaptive, real‑time PCR amplification using the icon96™ thermocycler with AutoNorm™ technology enables highly sensitive sequencing of cell‑free DNA (cfDNA) from ultra‑low input samples. Developed in collaboration with the Dana‑Farber Cancer Institute, the workflow demonstrates uniform library yields across variable inputs, improved on‑target capture efficiency, and reliable detection of variants down to 0.5% allele frequency, even from just 1 ng of cfDNA.

This application note demonstrates how iconPCR™ technology with AutoNorm™ on the icon96 real-time thermocycler enables balanced sequencing pools by dynamically controlling PCR amplification in each well. Unlike conventional fixed-cycle PCR workflows that require post-PCR quantification and normalization, AutoNorm monitors fluorescence in real time and stops amplification once a target threshold is reached. This automated normalization produces consistent library yields across variable inputs, simplifies NGS library preparation, reduces hands-on time by up to 60%, minimizes failed libraries, and maintains balanced read distribution for whole genome sequencing.

Design of Experiments (DOE) can dramatically improve PCR assay development, but traditional thermocyclers limit flexibility and efficiency. The icon96™ platform overcomes these limitations with 96 independently controlled wells, enabling multiple thermocycling conditions to be tested in a single plate. This technical note demonstrates how icon96 simplifies DOE workflows, reduces experimental runs, and accelerates assay optimization while minimizing time and consumable usage.

Explore how Revvity’s NEXTFLEX Small RNA-Seq Kit v4, combined with n6’s iconPCR system and AutoNorm technology, enables adaptive, real-time normalization of small RNA libraries from diverse human tissues, reducing PCR cycles and variability in library yield without altering RNA composition profiles.

16S rRNA gene sequencing is essential for exploring microbial diversity in various ecosystems, including soil, water, and human associated microbiomes. However, conventional PCR methods used in 16S library preparation often introduce significant biases due to overamplification, under-amplification, and the generation of chimeric sequences.

Discover how the icon96™, developed by n6, represents a transformative shift in PCR technology. Summarized in this comparative study, learn how using icon96 with iconPCR and conventional PCR workflows can produce high data quality, taxonomic resolution, and workflow consistency, especially when paired with long-read platforms like PacBio for full-length amplicon sequencing.

Single‑cell workflows are highly sensitive to sample quality, handling steps, and instrument settings, making it challenging to generate reliable, biologically meaningful data. Variability introduced during dissociation, staining, or capture, can reduce cell viability, skew population representation, and compromise downstream analysis.

This resource explores practical strategies to optimize each stage of single‑cell preparation, helping researchers improve sample integrity, reduce technical noise, and increase data quality. Learn how to gain clear guidance to streamline workflows, strengthen reproducibility, and achieve more confident single‑cell insights.

Standard PCR systems require users to set a predetermined number of cycles, based on assay and sample-specific factors. For single cell sequencing experiments, this requires knowledge of how many cells are captured and the RNA content within the cells. Different cell numbers and RNA quality require careful curation of the PCR cycle numbers for each sample, necessitating multiple thermocycler runs, which increases workflow complexity and potential sources of error.

Discover how the icon96™, when used together with AutoNorm™ can monitor each individual PCR reaction and terminate in real time based on fluorescence thresholds. This eliminates the guesswork of traditional workflows and ensures every library is amplified optimally.

n6 Tec and NEB offer a streamlined FFPE DNA library preparation workflow that enhances amplification consistency, reduces hands-on time by up to 60%, and maintains high data quality across variable inputs.

This application note highlights an optimized workflow using NEBNext UltraShear and NEBNext Multiplex Oligos with iconPCR AutoNorm, which dynamically tunes PCR cycling, minimizes batching and QC burden, and delivers uniform, high-quality NGS libraries from challenging FFPE samples.

Wednesday, March 25 at 15:00 GMT | 16:00 CET | 11:00 EDT | 8:00 PDT

What if your PCR step could adapt to every sample, instead of forcing every sample to adapt to your PCR?

In this now-on-demand webinar, Zach Herbert from Dana-Farber Cancer Institute's Molecular Biology Core Facilities shares how his team developed a targeted cfDNA sequencing workflow capable of detecting variants at <0.5% allele frequency from as little as 1 ng input — equivalent to just ~300 genome equivalents.

Watch this SelectScience® webinar to see real data from a genomics core lab:

• Library yield CV dropped from 54% to 8% (10 ng input) — 6x more uniform than fixed-cycle PCR
• On-target rates jumped from ~70% to >90% after implementing sample-specific amplification, across 1 ng and 10 ng inputs
• ~100 patient-derived cfDNA samples (2–25 ng range) processed in a single run, delivering balanced read counts (50–100M reads/pool) for hybrid capture
• Mean target coverage >200x at 1 ng input, with reliable variant detection down to 0.5% VAF using duplex sequencing

The challenge solved: cfDNA workflows are molecule-limited. Fixed-cycle PCR either over-amplifies (artifacts, distorted duplex families) or under-amplifies (insufficient yield for capture). The result? Imbalanced pools, wasted sequencing budget, and compromised sensitivity for rare variants.

The solution: Real-time, per-well PCR control that terminates amplification at each sample's optimal endpoint, preserving molecules, ensuring sufficient yield, and enabling mixed-input runs (1 ng + 10 ng together).

Why this matters for your lab:
"icon96 is really a game changer — shining a light into that black box [of PCR]. We can now see exactly the amplification path of every sample."
— Zach Herbert, Dana-Farber Cancer Institute

90%+ on-target isn't a benchmark. It's what Dana-Farber achieved, and what your cfDNA workflow can achieve.

Certificate of attendance
If you attend the live webinar, you will automatically receive a certificate of attendance, including a learning outcomes summary, for continuing education purposes.

If you view the on-demand webinar, you can request a certificate of attendance by emailing editor@selectscience.net.

Webinar details

• Cost: Free to attend
• Location: Online
• Duration: 60 minutes

Registration is required to secure your place. If you register but can’t attend live, you will receive a link to the on‑demand recording once it becomes available.

One of the biggest challenges in scaling genomics is maintaining data quality as throughput increases. As applications like single-cell analysis, spatial biology, MRD, cancer diagnostics, and microbiome research become more complex, traditional PCR and normalization workflows are struggling to keep up. In this SelectScience interview, Pranav Patel, Founder and CEO of n6 Tec, explains how the team is tackling this challenge with icon96™, the company's flagship amplification technology. By intelligently adjusting amplification based on sample quality and quantity, icon96 preserves data integrity, improves reproducibility, and eliminates the need for brute-force normalization.

This video was filmed at SLAS 2026.