Multiplex ligase-based genotyping methods combined with CE

In this genomic era, the ability to assay multiple genomic hot spots that have strong clinical implications is greatly desired. Conventional PCR‐based methods suffer from frequent false‐positive detections, particularly when a multiplex analysis is desirable. As an alternative to the error‐prone conventional methods, multiplex ligase‐based genotyping methods combined with CE have a strong potential. In this review, both previously developed methods and emerging methods are described to reveal the specificity, sensitivity, and simplicity of the ligase‐based methods. For each step (ligation, amplification, and separation), the principles of several alternative methods are discussed along with their applications to explore the future development of ligase‐based diagnostic methods.