![]() ![]() To evaluate the performance of Ultiplex, 294 out of 295 (99.7%) target primers were successfully designed. Ultiplex was developed as a free online multiplex primer design tool with a user-friendly web-based interface ( ). Thus, a user-friendly, highly automated and highly user-defined web-based multiplex PCR primer design software is needed to minimize the work of primer design and experimental verification. To conduct a multiplex PCR with higher than 100 plex multiplicity, primers need to be carefully designed to avoid the formation of secondary structures and nonspecific amplification between primers, templates and products. A high multiplicity PCR will outperform other technologies because of its lower cost, reaction time and sample consumption. A large number of variants have been employed in various medical applications, such as providing medication instructions, disease susceptibility testing, paternity testing, and tumour diagnosis.
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