Indigo dyeing is an ancient and worldwide natural dyeing technique that uses indigo dye (natural dye) as a dye material. Compared with synthetic dyes, natural dyes have better environmental affinity and no negative environmental and health issues. Taiwan’s indigo dyeing has a long history, and the Dutch introduced Indigofera tinctoria in the mid-17th century. After more than three centuries of historical evolution, Taiwan’s indigo dyeing has existed in Taiwan for nearly 400 years until the beginning of the 21st century. Taiwan’s indigo dyeing is constructed by indigo production process, indigo reduction process and dyeing process. Indigo production process and indigo reduction process are defined from the perspective of modern science, they are “biotechnology”, but the efficiency of this ancient biotechnology is not good. This study is based on the premise of environmental sustainability and human health, and constructs the quantitative operation procedures of indigo production process and indigo reduction process of Taiwan’s indigo dyeing to improve the preparation efficiency. This is not only conducive to the sustainable development of the human living environment, but also create opportunities to enhance the application value of Hakka culture and promote the development of indigo dyeing–related industries.