Antibodies specific to glycans are essential in many areas for many important fields, including disease diagnostics, therapeutics, and fundamental researches. However, due to their low immunogenicity and poor availability, glycans pose serious challenges to antibody development. Although molecular imprinting has developed into important methodology for creating antibody mimics with low cost and better stability, glycan-specific molecularly imprinted polymers (MIPs) still remain rather rare. Herein, we report a new strategy, precision imprinting with alternative templates, for the facile preparation of glycan-specific MIPs. Glycopeptides with desirable peptide length immobilized on a boronate affinity substrate were first prepared as alternative templates through in situ dual enzymatic digestion. A thinlayer was then produced to cover the glycans to an appropriate thickness through precision imprinting. With glycoproteins containing only N-glycans as well as both N- and O-glycans as glycan source, this approach was proved to be widely applicable and efficient. The strategy is particularly significant for the recognition of O-glycans, because enzymes that can release O-glycans from O-linked glycoproteins are lacking. The MIPs exhibited excellent glycan specificity. Specific extraction of glycopeptides and glycoproteins containing certain glycans from complex samples was demonstrated. This strategy opened a new avenue for the facile preparation of glycan-specific MIPs, facilitating glycan-related applications and research.