Dharma Valley Buddhist Temple is an important Buddhist site in Khotan area of Xinjiang. It boasts a high value of Buddhism and art, having presented a significant influence on Buddhist arts of China and even East Asia. In order to study the pigments and workmanship of wall paintings excavated from Dharma Valley Buddhist Temple ruins in Xinjiang, cultural relic fragments that cannot be restored were specially selected as samples. Wall painting samples were analyzed by stereomicroscopy, X-ray fluorescence spectrometry, X-ray diffractometry, Raman spectrometry and polarized light microscopy. The results show that the blue pigment is lazurite, the green pigment is atacamite, the white pigment is gypsum, the red pigment is cinnabar, the brown pigment is iron red and the ochre yellow pigment is ferrite yellow. This study provides an important scientific basis for the further exploration of the history of ancient arts and crafts and subsequent restoration and conservation.