The invention belongs to the technical field of biology and relates to a method for acquiring MYC genes in a human tissue by virtue of a two-step overlapping PCR method. The method comprises the following steps: (1) carrying out TES water bath dewaxing; (2) extracting DNA of a human tissue sample; (3) finding and selecting 7 MYC gene sequences with the same length; (4) designing specific primers of 16 MYC genes; (5) carrying out two-step overlapping PCR amplification; (6) purifying DNA segments; (7) detecting DNA concentrations; and (8) carrying out gene detection. The method is high in accuracy and efficiency, simple to operate and low in cost, the dewaxing effect is relatively good, andthe injure caused to the environment and a human body is relatively small; the DNA segments of 5279bp MYC target genes with different sequences, and the purities and concentrations of the target genes are relatively high; and segments of the target genes are verified by virtue of a sequencing method, and convenience is provided for the research of the biomedicine.