Novel Target Capture DNA Library Preparation Method Using CircLigase-Mediated Hook Ligation

Target capture DNA library preparation methods primarily use the biotin-streptavidin interaction to enrich target DNA, and these methods require complex operations and a time-consuming workflow, such as a series of wash steps at regulated temperatures. Here we present a new method that we call hook ligation for target DNA library preparation using circLigase to capture target DNA. We validated this concept by library preparation, sequencing, and acquiring the target DNA fragment information through bioinformatic analysis. We also provide an efficient reference point for single-strand DNA ligation to a hook sequence using circLigase, and we show that the efficiency can be influenced by the length and the position of the complementary area on the target DNA.