Effective Miniaturization of Illumina Nextera XT Library Prep for Multiplexed Whole Genome Sequencing and Microbiome Applications

The cost of sequencing per nucleotide has been declining over the past ten years; however library preparation of samples for next-generation sequencing remains a costly component of the process. In synthetic biology, microbial whole genome sequencing is becoming a routine process to validate genome editing, deconvolute mutagenesis campaigns, confirm high-throughput DNA construction, and survey microbiome samples. This application note details a method to miniaturize reaction volumes utilized in the Illumina Nextera XT DNA library kit, thereby reducing the overall cost per reaction while maintaining equivalent data to the standard protocol.