中国蔬菜Issue(7)：26-32,7.DOI:10.19928/j.cnki.1000-6346.2024.5021
单粒蔬菜种子DNA快速提取方法的建立
Establishment of Method for Quick Extraction of DNA from Single Vegetable Seed
摘要
Abstract
DNA was extracted from single seed as well as seedlings of seven vegetables,including of cabbage,Chinese cabbage,mustard,onion,cucumber,tomato and pepper using magnetic bead method.The seeds of these seven vegetables varied in size and shape.The DNA quality was evaluated by real-time fluorescent PCR reaction using reference gene Actin.In order to check whether the DNA quality was good enough for KASP(kompetitive allele specific PCR)assay,we conducted KASP assay for each vegetable.The results showed that the average Ct value of DNA extracted from single vegetable seed by magnetic beads was in the range of 19-24.KASP assays further confirmed their quality was good enough for KASP markers assay.In conclusion,this study established a simple,economical and practical method for DNA extraction of single seeds,which has good stability for vegetable seeds of different sizes and greatly reduces the time for obtaining genotype data.关键词
蔬菜种子/磁珠法DNA提取/KASP分子标记/实时荧光定量PCRKey words
vegetable seed/magnetic bead DNA extraction/KASP marker/real-time fluorescence quantitative PCR引用本文复制引用
杨帆,武剑,常立春,张涛,朱海峰,梁建丽,高杰,王晓武..单粒蔬菜种子DNA快速提取方法的建立[J].中国蔬菜,2024,(7):26-32,7.基金项目
财政部和农业农村部:国家现代农业产业技术体系项目(CARS-10-P19),新疆维吾尔自治区园艺学重点学科建设项目(2016-10758-37),中国农业科学院基本科研业务费专项(Y2023PT16),国家重点研发计划项目(2022YFF1003003) （CARS-10-P19）
