An object is to provide a method of manufacturing a liquid crystal layer in which an alignment film is repeatedly used and a liquid crystal compound in a liquid crystal layer can be sufficiently aligned. The method of manufacturing a liquid crystal layer includes: an alignment film forming step of forming an alignment film on a support; a liquid crystal alignment layer alignment step of laminating a first liquid crystal composition including a polymerizable liquid crystal compound on the alignment film and aligning the first liquid crystal composition; a liquid crystal alignment layer forming step of polymerizing the aligned first liquid crystal composition to form a liquid crystal alignment layer; a liquid crystal layer alignment step of laminating a second liquid crystal composition including a polymerizable liquid crystal compound on a surface of the liquid crystal alignment layer opposite to the alignment film and aligning the second liquid crystal composition; a liquid crystal layer forming step of polymerizing the aligned second liquid crystal composition to form a liquid crystal layer; and a liquid crystal layer separation step of separating the formed liquid crystal layer from the liquid crystal alignment layer, in which the liquid crystal layer alignment step to the liquid crystal layer separation step are repeated to repeatedly prepare the liquid crystal layer.