Abstract: In this study, twenty-two Agaricus bisporus monokaryons strains were pairwisely paired.Based on strain 9608, two different mating types of monokaryons were obtained.A + strains:9601, 9608, 9609, 9612, AgQG841-1, AgLH830-4, AgLH830-6, AgQL8125-5.A-:9602, 9603, 9604, 9607, Ag2k811-1, M7206-1, M7206-2, M7206-11, M7206-13.The genomic DNA of two strains were extracted and DNA library were constructed.The library was verified by restriction enzyme digestion with SalⅠ.The 20 kb, 14 kb and 9 kb 3 fragments were cut out, of which 20 kb and 9 kb were the inserted into the left and right arms of lambda vector and the fragment of 14kb, was the insertion genomic DNA fragments.Enzyme digestion result was in line with expectations, indicating the higher quality of the library.