Establishment of Method for Quick Extraction of DNA from Single Vegetable Seed
YANG Fan1,2,WU Jian2,CHANG Lichun2,ZHANG Tao1,2,ZHU Haifeng2,3,LIANG Jianli2,GAO Jie1*,WANG Xiaowu2*
1College of Horticulture,Xinjiang Agricultural University,Xinjiang Characteristic Horticultural Crop Germplasm Resources and Efficient Production Laboratory,Urumqi 830052,Xinjiang,China;2Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,State Key Laboratory of Vegetable Biobreeding,Beijing 100081,China;3Shanxi Agricultural University,Jinzhong 030801,Shanxi,China
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 goodenough 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.
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, 1(7): 26-32.
YANG Fan,WU Jian,CHANG Lichun,ZHANG Tao,ZHU Haifeng,LIANG Jianli,GAO Jie,WANG Xiaowu. Establishment of Method for Quick Extraction of DNA from Single Vegetable Seed. China Vegetables, 2024, 1(7): 26-32.