The present invention discloses a preparation method of scribing rubber cement of a tire tread. The scribing rubber cement of the tire tread is prepared from the following raw materials in parts by weight: 100 parts of natural rubber; 3.5 to 4.5 parts of zinc oxide; 1.5 to 2.5 parts of stearic acid; 15 to 25 parts of titanium white; 2.5 to 3.5 parts of insoluble sulfur; 0.9 to 1.1 parts of accelerant DM; 0.9 to 1.1 parts of rosin; and 15 to 25 parts of color masterbatch. The preparation method of the scribing rubber cement comprises the following steps: step 1) preparing rubber compound; and step 2) preparing the scribing rubber cement. The scribing rubber cement of the tire tread prepared by the present invention has high aesthetic degree and does not influence the product quality.