Branched oligosaccharide lycotetraose, β-D-glucopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucopyranosyl-(1→4)-β-D-galactopyranose, is a key constituent of many steroidal saponins, including glycoalkaloid α-tomatine, which is involved in protection of plants from invading pathogens. A new synthesis of the methyl β-lycotetraoside (2) is described. Key steps of the synthesis include two successive glycosylation reactions of disaccharide acceptor methyl (4,6-O-benzylidene-3-O-p-methoxybenzyl-β-D-glucopyranosyl)-(1→4)-2,3,6-tri-O-benzyl-β-D-galactopyranoside with readily available benzoylated trichloroacetimidates of α-D-glucopyranose and α,β-D-xylopyranose. This scheme allows sequential glycosylation in one-pot on account of the convenient in situ removal of a p-methoxybenzyl protecting group under the acid conditions of the first glycosylation step. Following deprotection, tetrasaccharide 2 was obtained in 19% yield over eight steps.