Construction method of amplicon sequencing library capable of reducing cross contamination

The invention discloses a construction method of an amplicon sequencing library capable of reducing cross contamination. The construction method comprises the following steps: obtaining sample DNA (Deoxyribose Nucleic Acid); performing first PCR amplification on the sample DNA by using the first primer, the second primer and the third primer to obtain an amplification product of the sample; performing purification; performing second PCR amplification on the purified product by using a fourth primer and a fifth primer to obtain an amplicon sequencing library; wherein the first primer sequentially comprises a first sequencing tag sequence and an amplicon specific forward primer from the 5'end to the 3 'end; the second primer sequentially comprises a second sequencing tag sequence and an amplicon specific reverse primer from the 5'end to the 3 'end; the third primer sequentially comprises a second sequencing linker sequence, a sample tag sequence and a second sequencing tag sequence from the 5'end to the 3 'end;