<FONT face="Verdana">水稻基因组DNA简易制备方法</FONT>

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,

doi:10.3969/j.issn.1001-7216.2010.06.021

中国水稻科学 >

2010 , Vol. 24 >Issue 6: 677 - 680

DOI: https://doi.org/10.3969/j.issn.1001-7216.2010.06.021

实验技术

水稻基因组DNA简易制备方法

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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

网络出版日期: 2010-11-10

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A Simple Method for Rapid Preparation of Rice Genomic DNA

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¹State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China; ²Agricultural College, Yangzhou University, Yangzhou 225009, China; ^#These authors contributed equally to this paper; *Corresponding authors, E-mail: qianqian188@hotmail.com; dalizeng@126.com

Received date: 1900-01-01

Revised date: 1900-01-01

Online published: 2010-11-10

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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（1~50 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

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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 . DOI: 10.3969/j.issn.1001-7216.2010.06.021

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

参考文献

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