Modelling and validating SiMSen-Seq Parameters for Precision DNA Sequencing

The overall aim of the project is to model and experimentally validate different theoretical parameters in the SiMSen-Seq library preparation method for optimal UMI-based sequencing accuracy and efficiency. Specifically, the study assessed the impact of variables such as the number of barcoding PCR cycles, reaction volumes in both barcoding PCR step, and the amount of barcoding PCR product transferred to the adapter PCR. Additionally, the project sought to estimate the number of unique molecular identifiers (UMIs) representing each genome copy to enhance precision in detecting low-frequency variants. Using human blood-derived genomic DNA (Roche), libraries were prepared and sequenced on an Illumina MiniSeq platform.