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The interaction between calf thymus DNA (ctDNA) and Pd (Ⅱ) meso-tetrakis(4-N-methyl pyridiniumyl) porphyrin (Pd-TMPyP) was investigated by steady solid-substrate room temperature phosphorescence (SS-RTP). The SS-RTP intensity and lifetime of Pd-TMPyP enhanced with the increasing ctDNA. The anion quenching experiment of Pd-TMPyP indicated that Pd-TMPyP intercalated into ctDNA, which was also approved by UV-Vis spectra.