iconPCR with AutoNorm™

PCR machines are at the forefront of the next-generation sequencing revolution. Traditional models apply the same conditions to all 96 samples, overlooking individual differences in input or amplification needs. n6's innovative technology revolutionizes this approach by allowing precise control over the cycling parameters at the level of each individual well. This breakthrough significantly enhances data quality, as demonstrated in a recent microbiome study. The study showed that n6's technology reduces PCR chimeras, achieves a more accurate representation of microbiomes, and streamlines sample normalization processes. These improvements not only lead to cost savings but also open up new possibilities for achieving superior data quality.

Every month we are analyzing thousands of clinical samples, typically from FFPE, of varying quality which influences library quality and yield. Many samples fail to meet our target amplification levels and require labor intensive manual intervention to prevent dropouts. The iconPCR technology potentially rescues previously failed or marginal libraries and tremendously simplifies the workflow allowing for near full automation of this crucial step and a lower overall number of failed tests. This is an exciting new technology that should find many uses in our laboratory.

iconPCR optimizes PCR cycles, the major user-defined step in NGS workflows. This overcomes sample handling bottlenecks and improves data quality. The auto-normalized libraries minimize under and over amplification, enhance data quality and improve experimental results.

As a product development scientist at 10x Genomics, I work to optimize the biochemical reactions in our products. Optimization of the times, temperatures, and number of cycles in each PCR is required to ensure highly reproducible results for our users. Traditionally, PCR cycle number in 10x workflows is dictated by cell number, RNA content, etc., and running substantially different samples and assays would require multiple thermal cyclers at once. My colleagues and I generated a set of Single Cell 3’ Gene Expression and Visium CytAssist Spatial Gene Expression libraries, which required 12 traditional thermal cyclers. In parallel, we generated the same libraries on a single run of N6 iconPCR, which dramatically streamlined the workflow. We used the auto normalization feature, which automatically halted amplification in individual wells at a set threshold above baseline. Sequencing the libraries yielded very comparable data, and I would not have been able to tell from which instrument the libraries were generated. Based on our initial evaluation, I'm excited for the release of the N6 iconPCR. (*N6 iconPCR is not a currently supported thermal cycler for use in 10x Genomics workflows.)