Miniaturized library preparation and large adapter sets for fast, cost-effective and high-throughput Next Generation Sequencing

Next-generation sequencing is an incredible tool for the discovery and validation of biomarkers, understanding the biology of diseases, assisting protein and antibody engineering, metagenomics and many more applications. While the sequencing cost per base is significantly decreased by high throughput sequencer, library preparation cost becomes a larger portion of the total costs. Additionally, maximizing the sample number on each flow cell and balance of data usage of each sample during multiplexed sequencing, which is often limited by the number of Unique Dual Index(UDI) adapters and sample normalization, also significantly affect the cost of NGS.
Here we used GenScript’s GenNature library preparation kit, a 2-step kit based on tagmentation, and compared different automation systems when handling miniaturized reaction volume, then adopted a protocol with the most reliable passing rate for low-depth sequencing. Utilizing our large UDI adapter sets, we demonstrated that we are able to sequence over 5000 samples each run, and reduce the total sequencing cost, including library preparation and sequencing cost by >80%. We also tested the limits of miniaturization library prep and large adapter sets for deep amplicon sequencing and showed even 1/2 volume can still be applicable for minimal depth over 1000x, with the average depth of 5800X on NextSeq550.