关键词: 大白菜, KASP, 反应体系优化

Abstract: In this study，the KASP（Kompetitive allele specific PCR）reaction system suitable for Chinese cabbage（Brassica rapa L. ssp. pekinensis）was established through optimization of three parameters（reaction volume，primer concentration and template DNA amount），which used genomic DNA from Chinese cabbage as template. The optimal KASP reaction system for 96-well plate format was as follows：total volume 8 μL，containing 1 μL template DNA（60–80 ng · μL-1），4 μL KASP Master mix （2×），0.14 μL primer assay mix（50 μmol · L-1）and 3 μL ddH2O. The optimal KASP reaction system for 384-well plate format was as follows：total volume 4 μL，containing 1 μL template DNA（60–80 ng · μL-1），2 μL KASP Master mix（2×），0.07 μL primer assay mix（50 μmol · L-1）and 1 μL ddH2O. With the optimal KASP reaction system，high quality results of KASP-SNP genotyping were produced on different Chinese cabbage accessions with different primers，which indicated the stability of the current system. The optimized KASP reaction system can be used in genetic study for Chinese cabbage，such as genetic diversity analysis，gene mapping and genetic map construction.

Key words: Chinese cabbage, KASP, reaction system optimization