The IAB company developed a robust WGS protocol including all essential QC steps, library preparation, and a very accurate and validated bioinformatics pipeline under the EN ISO 15189. For NovaSeq 6000 we established an evaluation method of NGS libraries equimolarity and pool loading concentration using pre-sequencing on iSeq. Presequencing on iSeq platform is not only suitable for the quality control of NGS libraries, but it can save time and money as well. Due to the implantation of all QC steps including pre-sequencing, our sequencing data are of very high quality with average read depth > 50X. The draft of the digital genome map, created from 100 samples of the Czech population, was generated and will be expanded to up to 1,000 samples.