5种提取方法对甘蔗汁中DNA提取的效果
Effects of Five Methods on Total DNA Extraction from Sugarcane Juice
曾巧英 1凌秋平 1胡斐 1刘睿1
作者信息
- 1. 广东省甘蔗改良与生物炼制重点实验室/广东省生物工程研究所(广州甘蔗糖业研究所),广东广州510316
- 折叠
摘要
[目的]研究5种提取方法对甘蔗汁中DNA提取的效果.[方法]以甘蔗种茎中获得的蔗汁为材料,采用2种植物基因组提取试剂盒柱式法和磁珠法、SDS法、CTAB法及简化提取法5种方法提取DNA,对比所提取的DNA浓度、纯度及后续PCR扩增的影响.[结果]以SDS法提取获得的蔗汁DNA浓度最高,平均可达377.50 ng/mL.而柱式法的试剂盒,提取的DNA浓度最低,PCR检测没有能够获得特异的目标条带,不适合用于蔗汁DNA的直接提取.虽然简化法提取的DNA浓度较低,但是能够满足PCR扩增的要求,由于不涉及氯仿等有害物质,操作简单,适用于对DNA质量要求不高时的快速提取.[结论]SDS法最适合用于蔗汁中DNA的提取.
Abstract
[Objective]To study the effects of five methods on total DNA extraction from sugarcane juice.[Method]In this study, five meth-ods, including two kinds of the plant DNA extraction kit (with column and magnetic bead, respectively), CTAB method, SDS method and simplified DNA extraction method were used to extract DNA from sugarcane juice collected from seed-stem, and the quality and quantity of DNA and the following PCR analysis were compared.[ Result] SDS method was appropriate for DNA extraction from sugarcane juice and ob-tained highest amount of DNA from sugarcane juice (377.50 ng/μL, on average).DNA yield from the plant DNA extraction kit (with col-umn) was lowest and might interfere the following PCR analysis, so this method was not suitable for directly extracting DNA from sugarcane juice.The simplified DNA extraction method had advantage of simple and fast.Although its yield and purity were lower than those extracted by SDS and CTAB methods, it meet the demand of the following PCR analysis and was suitable for fast extraction with lower quality requirement. [Conclusion] SDS method was appropriate for DNA extraction from sugarcane juice.
关键词
甘蔗/蔗汁/DNA提取Key words
Sugarcane/Sugarcane juice/DNA extraction引用本文复制引用
基金项目
国家甘蔗产业技术体系(CARS-20-1-4)
广东省科技计划(2014A040401033)
广东省科学院科研平台环境与能力建设项目(2016GDASPT-0306)
出版年
2017
国家科技期刊平台
NSTL
维普
万方数据