福建农业科技2023，Vol.54Issue(10)：40-47,8.DOI:10.13651/j.cnki.fjnykj.2023.10.007
一种适用于高通量基因分型的水稻种子切片DNA制备技术
Preparation Technique of DNA Extracted from the Slices of Rice Seeds Suitable for the High-throughput Genotyping
摘要
Abstract
The preparation method of DNA with high quality and low cost extracted from the slices of rice seeds was established for the high-throughput genotyping of breeding populations.The degree of purity and concentration of DNA extracted from the leaves of rice by TPS method and those of DNA extracted from the slices of rice seeds by u-sing the method of 96-well plates combined with magnetic beads were compared in this paper.Then,the quality of DNA extracted from the slices of rice seeds by using the method of 96-well plates combined with magnetic beads was further verified by ST ARP(Semi-thermal asymmetric PCR)marker detection.The results showed that the quality and the degree of purity of DNA extracted from the slices of rice seeds by using the method of 96-well plates combined with magnetic beads were better than those of DNA extracted from the leaves of rice by TPS method.The verification results of STARP marker detection showed that the method of 96-well plates combined with magnetic beads to extract the DNA from the slices of rice seeds could be applied to the STARP marker genotyping of GS3 and Wx,and the identification results were consistent with the sequencing results.The results showed that the method of 96-well plates combined with magnetic beads could be used to extract the DNA with high recovery rate,high purity,high in-tegrity and no PCR inhibitors from the trace slices of rice seeds for the analysis of gene haplotype with the high-throughput STATP method.Therefore,the operation flow of high-throughput genotyping for the DNA extraction by using the method of 96-well plates combined with magnetic beads could be obtained.关键词
水稻种子切片/96孔板结合磁珠法/高通量基因分型/STARP技术Key words
Slices of rice seeds/Method of 96-well plates combined with magnetic beads/High-throughput genoty-ping/STARP(Semi-thermal asymmetric PCR)technology分类
农业科技引用本文复制引用
林旻,杨绍华,林艳,王月,胡太蛟,宋亚娜,潘雷博,陈在杰..一种适用于高通量基因分型的水稻种子切片DNA制备技术[J].福建农业科技,2023,54(10):40-47,8.基金项目
福建省科技计划公益类项目(2020R1027009) （2020R1027009）
福建省人民政府、中国农业科学院"5511"协同创新工程(XTCXGC2021002). （XTCXGC2021002）
