| 水稻基因组DNA简易制备方法 |
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| 引用本文： | 孙川,陈刚,饶玉春,张光恒,高振宇,刘坚,鞠培娜,胡江,郭龙彪,钱前,曾大力.水稻基因组DNA简易制备方法[J].中国水稻科学,2010,24(6):677-680. |
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| 作者姓名： | 孙川 陈刚 饶玉春 张光恒 高振宇 刘坚 鞠培娜 胡江 郭龙彪 钱前 曾大力 |
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| 作者单位： | 1. 中国水稻研究所,水稻生物学国家重点实验室,浙江,杭州,310006;扬州大学,农学院,江苏,扬州,225009
2. 中国水稻研究所,水稻生物学国家重点实验室,浙江,杭州,310006 |
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| 基金项目： | 国家转基因生物新品种培育重大科技专项资助项目，国家自然科学基金，浙江省自然科学基金杰出青年团队项目 |
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| 摘 要： | 介绍了一种用于PCR的水稻基因组DNA的快速制备方法。该方法只需将少量(1～50mg)样品、适量提取液(500μL)和一粒钢珠放入2mL离心管,在细胞破碎仪中粉碎2min,经离心后可直接取少量(约5μL)上清液作为PCR的模板DNA,也可进一步根据需要实现高质量DNA的提取。该方法具有成本低、操作快速简便等优点,一个人可以在10min内完成96份样品DNA的简易制备,特别适合高通量的分子检测。
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| 关 键 词： | DNA提取 高通量PCR扩增 样品制备 方法 水稻 |
| 收稿时间： | 1900-01-01; |
A Simple Method for Rapid Preparation of Rice Genomic DNA |
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| SUN Chuan,CHEN Gang,RAO Yu-chun,ZHANG Guang-heng,GAO Zhen-yu,LIU Jian,JU Pei-na,HU Jiang,GUO Long-biao,QIAN Qian,ZENG Da-li.A Simple Method for Rapid Preparation of Rice Genomic DNA[J].Chinese Journal of Rice Science,2010,24(6):677-680. |
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| Authors: | SUN Chuan CHEN Gang RAO Yu-chun ZHANG Guang-heng GAO Zhen-yu LIU Jian JU Pei-na HU Jiang GUO Long-biao QIAN Qian ZENG Da-li |
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| Institution: | State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China; Agricultural College, Yangzhou University, Yangzhou 225009, China; These authors contributed equally to this paper; *Corresponding authors, E-mail: qianqian188@hotmail.com; dalizeng@126.com |
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| Abstract: | A simple method for preparation of rice genomic DNA was developed. A small amount (1- 50 mg) of leaf tissue of rice seedling, 500 μL of extraction buffer, and one steel bead were put into a 2 mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of the supernatant was directly applied to PCR amplification. Otherwise, the supernatant was precipitated with 2 volumes of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 minutes. It is especially suitable for genotyping of large number of samples. |
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| Keywords: | DNA extraction high-throughput PCR amplification sample preparation methodology rice |
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