During the period of 2011 to 2013, a wall painting fragment was unearthed from affiliated building remains in Emperor Qinshihuang’s Mausoleum near Emperor Qinshihuang’s Mausoleum Site Museum. In order to understand the techniques and materials of the wall painting, polarized light microscopy (PLM) , Raman microscopy (RM), scanning electron microscopy and energy dispersive X-ray spectrometry (SEM-EDS), infrared spectrometry (IR) and X-ray diffraction (XRD) were utilized to analyze the pigments and the painting techniques. The results show that the wall painting is composed of a pigment layer, a finely ground plaster layer and a coarse plaster layer. The white ground layer is calcium carbonate limestone, the black pigment is black copper (melaconite) and the red outlining pigment is hematite. There is calcium carbonate in the plaster layer and plant stalk in the coarse plaster layer. This is the first time when a wall painting fragment was unearthed at Emperor Qinshihuang’s Mausoleum and this work is the first study of a wall painting of Emperor Qinshihuang’s Mausoleum. This study supplies important information about Emperor Qinshihuang’s Mausoleum and its affiliated building.