深圳大学学报理工版

李琳,罗淋淋,罗光宇,等.一种植物基因组DNA快速提取方法的建立与评估[J].深圳大学学报理工版,2020,37(1):1-8.[doi:10.3724/SP.J.1249.2020.01001]
LI Lin,LUO Linlin,LUO Guangyu,et al.Establishment and evaluation of a method for rapid extraction of plant genomic DNA[J].Journal of Shenzhen University Science and Engineering,2020,37(1):1-8.[doi:10.3724/SP.J.1249.2020.01001]

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一种植物基因组DNA快速提取方法的建立与评估

李琳^1,2,罗淋淋^1,2,3,罗光宇^1,2,3,莫蓓莘^1,2,刘琳^1,2

1)深圳大学生命与海洋科学学院,广东省植物表观遗传学重点实验室,广东深圳518060; 2)深圳大学龙华生物产业创新研究院,广东深圳518060; 3)深圳大学物理与光电工程学院,广东深圳518060

关键词：分子生物学; 基因组脱氧核糖核酸; 快速提取; 聚合酶链式反应; 水稻; 拟南芥; 基因型鉴定; 单核苷酸多态性分析; 简单重复序列标记分析

Establishment and evaluation of a method for rapid extraction of plant genomic DNA

LI Lin^{1, 2}, LUO Linlin^{1, 2, 3}, LUO Guangyu^{1, 2, 3}, MO Beixin^{1, 2}, and LIU Lin^{1, 2}

1)College of Life Sciences and Oceanography, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China2)Longhua Bioindustry and Innovation Research Institute, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China3)College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China

Keywords： molecular biology; genomic deoxyribonucleic acid; rapid extraction; polymerase chain reaction; Oryza sativa L.; Arabidopsis thaliana; genotyping; single nucleotide polymorphisms analysis; simple sequence repeat analysis

DOI: 10.3724/SP.J.1249.2020.01001

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植物基因组脱氧核糖核酸(deoxyribonucleic acid, DNA)的提取是植物分子实验的基础,但随着样本数量的增加,基因组DNA的提取成为植株基因型鉴定以及单核苷酸多态性检测等分子实验的限速环节.为解决现有技术中提取植物基因组DNA过程繁琐、耗时耗力的问题,从提取液配方和提取条件等方面改良常规快速植物基因组DNA提取法,缩短了样本制备时间并简化了提取步骤,改良后每个样本DNA制备需时不到90 s,同时采用96孔板和配套设施显著提高了实验通量.该方法对植物样本的状态要求低、扩增效率高、成本低,适用于拟南芥、水稻和玉米等多种植物基因组DNA的提取与聚合酶链式反应(polymerase chain reaction, PCR)扩增实验,尤其适用于大样本量的规模化基因型鉴定.

The extraction of plant genomic deoxyribonucleic acid(DNA)is the basis for plant molecular experiments. However, as the number of plant samples increases, genomic DNA extraction becomes the rate-limiting step in molecular experiments such as the identification of plant genotypes and single nucleotide polymorphisms detection. Regular DNA extraction is usually complicated and time-consuming and effort-consuming. To cope with these problems, we improve the conventional rapid plant genomic DNA extraction method from the aspects of extraction buffer and extraction conditions to further shorten the sample preparation time and simplify the extraction steps. After optimization, the preparation of each DNA sample takes less than 90 s, and adoption of 96 wells plates and related facilities significantly improve experimental throughput. This system has low requirements for the status of plant samples, and has high amplification efficiency at low cost. Besides, this system is suitable for a variety of plant species such as Arabidopsis thaliana L., rice(Oryza sativa L.), and maize(Zea mays L.)for genomic DNA extraction and polymerase chain reaction(PCR)amplification, especially for genotyping experiments that require a large number of samples.

引言
1 材料与方法
2 结果
3 结论和讨论

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