Tencel Jacquard fabric is one of the eco-fabrics used for underwear, sportswear, bedclothes, clothes for aged people, and hospital textiles. It is popular for these uses because it is easy to process into yarns and fabrics alone or in blends, very stable in washing and drying, thermally stable, and easy to dye with deep vibrant colors using direct, reactive, or vat dyes. In order to provide antimicrobial properties for Tencel Jacquard fabrics, they were treated with ginkgo biloba extract and silicon softener using two different processes so the results could be compared. One of the processes treated the fabrics with ginkgo biloba extract and silicon softener simultaneously, and the other process treated the fabrics with these agents sequentially. The treated Tencel Jacquard fabrics were characterized by scanning electron microscopy, and their antimicrobial properties were evaluated. In addition, water repellency, air permeability, water vapor permeability, and yellowness were measured. It was observed that the ginkgo biloba extract and silicon softener were present on the surface of the treated fibers, and the quantity of these agents before and after laundering was proportional to the measured antimicrobial activity of the fabrics before and after laundering. Fabrics treated with both agents had stronger water/oil repellency than fabrics treated with only ginkgo biloba extract. As the quantities of the two treating agents on fabrics were increased, their air permeability and water vapor permeability decreased. No significant changes were observed for yellowness based on the amounts of treating agents applied. From these results, it is expected that Tencel fabrics treated with ginkgo biloba extract and silicon softener are excellent for use as bedclothes.