The Quick-ITS Plus NGS Library Prep Kit (UDI) is the fastest and simplest NGS library prep targeting the ITS region for high-throughput sequencing, now with unique dual indexing for improved barcode resolution. The automation-friendly protocol utilizes a single qPCR/PCR for combined targeted amplification and barcode addition using specially designed primers. After pooling by equal volume, a single clean-up of the final library is performed, rather than massive AMPure® bead-based clean-ups. Additional library quantification analysis such as TapeStation® analysis or gel electrophoresis are not necessary. With these features, the workflow dramatically reduces the hands-on time of library preparation to only 30 minutes.