This research combined the self-ordered ring (SOR) technology with solid substrate-room temperature phosphorimetry (SS-RTP) and developed a new (ARS) n -P-SOR (ARS is alizarin red S, (ARS) n refers to multiple ARS molecules, P is piperidine). (ARS) n -P-SOR could increase the number of ARS molecules in the biological target, which could further improve the sensitivity of SS-RTP. Thus, a new SS-RTP for the determination of trace deoxyribonucleic acid (DNA) was established using (ARS) n -P-SOR as a phosphorescent probe. This method was successfully applied to the determination of DNA in nectar. Meanwhile, the mechanism of determining trace DNA by (ARS) n -P-SOR was discussed.