中国水稻科学

• 实验技术 • 上一篇    

水稻基因组DNA简易制备方法

孙川1,2,#; 陈刚1,2,#;饶玉春1;张光恒1;高振宇1;刘坚1;鞠培娜1;胡江1;郭龙彪1;钱前1,*;曾大力1,*   

  1. 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; 2扬州大学 农学院, 江苏 扬州 225009;#共同第一作者;*通讯联系人, E-mail: qianqian188@hotmail.com; dalizeng@126.com
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-11-10 发布日期:2010-11-10

A Simple Method for Rapid Preparation of Rice Genomic DNA

SUN Chuan1,2, #;CHEN Gang1,2, #; RAO Yu-chun1; ZHANG Guang-heng1;GAO Zhen-yu1; LIU Jian1; JU Pei-na1; HU Jiang1; GUO Long-biao1;QIAN Qian1,*;ZENG Da-li 1,*   

  1. 1 State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China; 2 Agricultural College, Yangzhou University, Yangzhou 225009, China; # These authors contributed equally to this paper; *Corresponding authors, E-mail: qianqian188@hotmail.com; dalizeng@126.com
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-11-10 Published:2010-11-10

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(1~50 mg)样品、适量提取液(500 μL)和一粒钢珠放入2 mL离心管,在细胞破碎仪中粉碎2 min,经离心后可直接取少量(约5 μL)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10 min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。

关键词: DNA提取, 高通量PCR扩增, 样品制备, 方法, 水稻

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.

Key words: DNA extraction, high-throughput PCR amplification, sample preparation, methodology, rice