表面活性剂LAS在棉花高通量DNA提取中的应用  

Application of LAS in High-throughput DNA Extraction from Cotton

作　　者：陈丽婷 黄龙雨 黄义文 吴玉珍 周大云 艾先涛[3] 彭军 吴立强[1] 匡猛 Chen Liting;Huang Longyu;Huang Yiwen;Wu Yuzhen;Zhou Dayun;Ai Xiantao;Peng Jun;Wu Liqiang;Kuang Mengg(Hebei Research Base for State Key Laboratory of Cotton Biology,College of Agronomy,Hebei Agricultural University,Baoding,071001;National Nanfan Research Institute(Sanya),Chinese Academy of Agricultural Sciences,Sanya,572024;Institute of Economic Crops,Xinjiang Academy of Agricultural Sciences,Urumqi,830091)

机构地区：[1]河北农业大学农学院,棉花生物学国家重点实验室河北基地,保定071001 [2]中国农业科学院国家南繁研究院,三亚572024 [3]新疆农业科学院经济作物研究所,乌鲁木齐830091

出　　处：《分子植物育种》2023年第16期5301-5308,共8页Molecular Plant Breeding

基　　金：中国农业科学院基本科研业务费专项院级统筹项目(Y2022XK21);国家棉花产业技术体系项目(CARS-15-26);新疆维吾尔自治区科技支疆项目计划(指令性)项目(2020E02097)共同资助。

摘　　要：本研究以棉花种子和叶片为材料,探索一种适用于棉花DNA高通量提取的方法。通过在裂解液中加入APG、LAS、CPB、DTAB、Tween80、SDS和CTAB 7种不同表面活性剂,比较其在棉花种子和叶片中的DNA提取效果,并与磁珠法和氯仿抽提法两种高通量纯化方法结合,对比传统SDS和CTAB提取液提取效果。结果表明,加入LAS表面活性剂的裂解液提取的DNA效果较好,提取棉花种子和叶片的浓度可达2127.61和572.70 ng/μL,OD_(260/280)分别为1.97和1.98,上清液清澈透明。LAS与两种纯化方式结合的DNA提取效果都明显优于传统CTAB和SDS提取液的提取效果,其中LAS与氯仿抽提的纯化方法结合提取效果最好,且DNA提取通量可达到600个样品/2 h。LAS表面活性剂能在棉花种子和叶片DNA提取中都能起到促进作用,与氯仿抽抽提纯化方法相结合可兼顾DNA提取通量与质量,能满足后续大部分分子实验要求。In this study,cotton seeds and leaves were used as materials to explore a high-throughput DNA extraction method for cotton.By adding seven different surfactants APG,LAS,CPB,DTAB,Tween80,SDS and CTAB to the lysate,the DNA extraction effects of APG,Las,CPB,SDS and CTAB in cotton seeds and leaves were compared,and the extraction effects of traditional SDS and CTAB extracts were compared with those of magnetic beads and chloroform extraction.The results showed that the lysate with LAS surfactant had a good effect on DNA extraction,and the concentration of extracted cotton seeds and leaves could reach 2127.61 and 572.70 ng/μL.OD_(260/280) were 1.97 and 1.98 respectively,and the supernatant was clear and transparent.The DNA extraction effect of Las combined with the two purification methods is significantly better than that of traditional CTAB and SDS extraction solutions.The combination of LAS and chloroform extraction has the best extraction effect,and the DNA extraction flux can reach 600 samples/2 hour.Las surfactant can promote DNA extraction from cotton seeds and leaves.Combined with chloroform extraction and purification method,it can give consideration to DNA extraction flux and quality,and can meet the requirements of most subsequent molecular experiments.

关 键 词：棉花 LAS DNA提取 表面活性剂 高通量 

分 类 号：S562[农业科学—作物学]