One-pot sequential glycosylations are unambiguously the most straightforward strategies for the construction of complex glycans. Among them, sequential activation of thioglycosides provided one of the most powerful platforms. In this review, we summarized the developments, recent advances and synthetic utilizations of one-pot glycosylation relying on relative reactivity value (RRV) controlled successive activation of thioglycosides, pre-activation based iterative activation of thioglycosides and the glycosyl sulfoxide activation initiated sequential activation of thioglycosides.