Application Note: Effective Miniaturization of Illumina Nextera XT Library Prep for Multiplexed Whole Genome Sequencing and Microbiome Applications
27 February 2018
Reduce your sequencing costs through high-throughput miniaturized sequencing reaction volume process.
The cost of sequencing per nucleotide has been declining by orders of magnitude 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.
Here we detail 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. We then tested this miniaturized method on a diverse variety of microbes to show that the method is effective across samples. Finally, we applied this method to a microbiome study and found that it maintains sample diversity, ratios, and identification.