| 夏澳运，彭子艾，李丹丹，赵壮壮，王 慧，陈志强，郭 涛.一种适用于高通量基因分型的水稻 DNA样本制备技术研究[J].广东农业科学,2019,46(8):1-7 |
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一种适用于高通量基因分型的水稻 DNA样本制备技术研究 |
| An Extraction Technique of Rice DNA SamplesSuitable for High-throughput Genotyping |
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| DOI：10.16768/j.issn.1004-874X.2019.08.001 |
| 中文关键词: 水稻 根 96 孔板 DNA 提取 高通量 |
| 英文关键词: rice root 96-orifice plate DNA extraction high-throughput |
| 基金项目:国家重点研发计划项目（2016YFD0102102）；广东省科技计划项目（2018B020206002）；国家现代农业产业技术体系建设专项（CARS-01-12） |
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| 中文摘要: |
| 【目的】提供一种基于 96 孔板的水稻根部 DNA 提取技术，可显著提升 DNA 提取效率。【方法】
在 96 孔深孔板的每个孔中放入一粒萌发的种子，在深孔板下方对应放置 96 孔 PCR 板，在适当光温下促使水稻
根伸入 PCR 板中；当水稻幼苗发育至一叶一心时，将 PCR 板移至干冰与 95% 乙醇混合液体中，利用冷冻处理
捕获幼根；用改良的 TRIS-EDTA 法批量提取 DNA 并进行浓度和纯度检测。【结果】从播种到根部组织样品的
获取全部在 96 孔板中进行，实现了样本组织的批量获取及目标个体的精确对应。改良的 TRIS-EDTA 法提取 96
个水稻样本 DNA 的效率显著高于传统的 CTAB 法。【结论】基于 96 孔板的水稻根部组织批量获取及 DNA 高效
提取方法，对于提升水稻分子育种效率具有实践意义。 |
| 英文摘要: |
| 【Objective】This paper provides a DNA extraction technique based on 96-orifice plates for rice roots,
which can significantly improve DNA extraction efficiency. 【Method】 A germinated seed was placed in each orifice of a 96
deep-orifice plate, and a 96-orifice PCR plate was placed under the deep orifice plate to promote the rice root stretching into
the PCR plate at an appropriate light and temperature. When the rice seedlings developed with two leaves, the PCR plate was
moved to a mixed liquid of dry ice and 95% ethanol, and the roots were captured by freezing treatment. DNA was extracted
in bulk by using the modified TRIS-EDTA method and the concentration and purity of DNA was detected . 【Result】 The
results showed that all the processes from the seeding to the obtaining of root tissue samples were carried out in 96-orifice
plates, which achieved the mass abtainment of the sample tissues and the accurate correspondence of the target individuals.
Compared with other methods, the efficiency of extracting DNA from 96 rice samples by the modified TRIS-EDTA method
was significantly higher than that of the traditional CTAB method. 【Conclusion】 The batch obtainment of rice root
tissues based on 96-orifice plate and DNA efficient extraction method reported in this paper has practical significance for
improving of rice molecular breeding efficiency. |
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