According to the results of previous related work, we prepare 95 different conditions for HTE. The details of the 95 conditions are shown in Table of the SI. It should be noted that all commercial coupling reagents were involved in conditions set except for acyl chloride, because it is not compatible with DMF solvent. With the conditions and substrates pairs in hand, we performed the HTE to collect reaction yield data. Our HTE workflow comprises several key steps: experiment design, high-throughput experimental preparation, high-throughput reaction setup, high-throughput reaction work-up, and high-throughput detection and analysis. The experimental design was tailored for HTE, like with careful consideration of well placement to minimize variability. Following this, automated systems were used for the precise preparation and setup of a large number of reactions in parallel.