Next-generation sequencing of highly multiplexed samples: the NuGEN Encore™ 384 Multiplex System
Advances in DNA sequencing throughput afforded by highly parallel, reversible-terminator sequencing platforms have completely redefined our view of biological inquiry. Crisp, high-resolution, digital views of whole-genome biology provide both profound clarity and significance. As with any disruptive technology, implementation creates turbulence. In the case of NGS, accelerated sequencing throughput has created challenges in upstream sample preparation and downstream data analysis. Initially, sample preparation was particularly problematic because it required individuals with highly specialized molecular biology skills for the hands-on execution of complex protocols. Fortunately, automated solutions for NGS library preparation have arrived, and it is reasonable to expect broad dissemination of turnkey solutions for routine library generation in the very near future. What remains problematic is that some sequencing projects involve large numbers of samples that require relatively shallow sequencing coverage. The sequencing costs of such projects are often prohibitive, calling for methods to multiplex high sample numbers in the same sequencing run. The obvious solution is to combine many samples into a single sequencing lane, and there are now many reports in the literature describing ad hoc methods and strategies for multiplexing samples prior to sequencing. Here we introduce a commercial solution for deep multiplexing of NGS samples that builds on our Encore™ NGS Multiplex System. The Encore 384 Multiplex System consists of a refined set of 384 molecularly “bar-coded” library adaptors that enable deep multiplexing of sequencing samples within a library preparation system that is compatible with high-throughput automation. The version of the product discussed in this White Paper contains adaptor sequences compatible with Illumina sequencing platforms.