石斛基因组DNA提取方法比较研究  

Comparison Study on Genomic DNA Extraction Methods for Dendrobe

作　　者：潘映秋 洪亮 卢启寰 李兆奎 夏慧丽 PAN Ying-qiu;HONG Liang;LU Qi-huan;LI Zhao-kui;XIA Hui-li(Taizhou Key Laboratory of Drug Composition and Adulteration Identification Technology,Taizhou Food Inspection and Testing Center/Taizhou Institute of Drug Inspection,Taizhou 318000,China)

机构地区：[1]台州市食品检验检测中心/台州市药品检验研究院、台州市药物成分与掺伪鉴定技术重点实验室,浙江台州318000

出　　处：《中国食物与营养》2023年第7期26-30,共5页Food and Nutrition in China

基　　金：浙江省市场监督管理局科研计划项目(项目编号:20210174)。

摘　　要：目的:比较3种基因组DNA提取试剂盒对提取富含多糖多酚类物质的铁皮石斛和霍山石斛鲜叶及冻叶的提取效果。方法:对提取的DNA进行纯度及产率检测、电泳质量分析及ITS2 PCR扩增效率检测。结果:市售基于蛋白酶K消化法的试剂盒不适用于石斛DNA提取;基于改良CTAB法的试剂盒更适用于需要较高DNA模板量的石斛鲜叶的DNA提取;基于改良SDS法的试剂盒更适用于石斛冻叶的DNA提取,该法提取的鲜叶DNA完整性最佳,更适用于对DNA完整度要求较高的DNA大片段的分析研究。结论:基于改良CTAB法和改良SDS法的试剂盒提取的石斛鲜叶及冻叶的基因组DNA均适用于常规分子检测。Objective To compare the effectiveness of three kinds of genomic DNA extraction kits for fresh and frozen leaves of Dendrobium officinale and Dendrobium huoshanense rich in polysaccharides and polyphenols.Method DNA purity and yield tests,DNA electrophoresis quality analysis,and ITS2 PCR amplification efficiency were analyzed.Result The commercially available kit based on proteinase K digestion was not suitable for dendrobe DNA extraction.The modified CTAB-based kit was more suitable for extracting DNA from fresh leaves requiring a higher template amount,while the modified SDS-based kit was more suitable for extracting DNA from frozen leaves,producing the highest quality fresh leaf DNA for experiments requiring high DNA integrity.Conclusion The kits based on the modified CTAB Method and SDS Method were both suitable for conventional molecular detection of dendrobe genomic DNA from fresh and frozen leaves.

关 键 词：石斛 DNA提取方法 SDS法 CTAB法 

分 类 号：R282.5[医药卫生—中药学]